Antibody data
- Antibody Data
- Antigen structure
- References [38]
- Comments [0]
- Validations
- Western blot [2]
- Other assay [32]
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Validation data
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- Product number
- MA5-15739-HRP - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- beta Actin Monoclonal Antibody (BA3R), HRP
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- MA5-15739-HRP has been successfully used in Western blotting applications with human, mouse, rat, rabbit, and chicken samples.
- Reactivity
- Human, Mouse, Rat, Canine, Chicken/Avian, Rabbit
- Host
- Mouse
- Conjugate
- Horseradish Peroxidase
- Isotype
- IgG
- Antibody clone number
- BA3R
- Vial size
- 50 µL
- Concentration
- 1 mg/mL
- Storage
- 4° C
Submitted references miR-340-5p inhibits pancreatic acinar cell inflammation and apoptosis via targeted inhibition of HMGB1.
Evidence for reprogramming of monocytes into reparative alveolar macrophages in vivo by targeting PDE4b.
Checkpoint inhibition through small molecule-induced internalization of programmed death-ligand 1.
Distinct mRNAs in Cancer Extracellular Vesicles Activate Angiogenesis and Alter Transcriptome of Vascular Endothelial Cells.
Inhalation of lung spheroid cell secretome and exosomes promotes lung repair in pulmonary fibrosis.
Beneficial Effects of Acetyl-DL-Leucine (ADLL) in a Mouse Model of Sandhoff Disease.
Transcutaneous Vagus Nerve Stimulation Regulates the Cholinergic Anti-inflammatory Pathway to Counteract 1, 2-Dimethylhydrazine Induced Colon Carcinogenesis in Albino wistar Rats.
Hydralazine targets cAMP-dependent protein kinase leading to sirtuin1/5 activation and lifespan extension in C. elegans.
Repurposing mechanistic insight of PDE-5 inhibitor in cancer chemoprevention through mitochondrial-oxidative stress intervention and blockade of DuCLOX signalling.
Activation of prolyl hydroxylase-2 for stabilization of mitochondrial stress along with simultaneous downregulation of HIF-1α/FASN in ER + breast cancer subtype.
KMT5A promotes metastasis of clear cell renal cell carcinoma through reducing cadherin-1 expression.
Hyperexcitability of the local cortical circuit in mouse models of tuberous sclerosis complex.
Developmental NMDA receptor dysregulation in the infantile neuronal ceroid lipofuscinosis mouse model.
DuCLOX-2/5 Inhibition Attenuates Inflammatory Response and Induces Mitochondrial Apoptosis for Mammary Gland Chemoprevention.
Ubiquitin-Mimicking Peptides Transfer Differentiates by E1 and E2 Enzymes.
Functional and structural characterization of a novel malignant hyperthermia-susceptible variant of DHPR-β(1a) subunit (CACNB1).
Selective Cannabinoid 2 Receptor Stimulation Reduces Tubular Epithelial Cell Damage after Renal Ischemia-Reperfusion Injury.
Estradiol signaling mediates gender difference in visceral adiposity via autophagy.
Evaluation of the effects of the combination of BMP-2-modified BMSCs and PRP on cartilage defects.
Disruption of mesoderm formation during cardiac differentiation due to developmental exposure to 13-cis-retinoic acid.
Human snoRNA-93 is processed into a microRNA-like RNA that promotes breast cancer cell invasion.
Alpha-linolenic acid stabilizes HIF-1 α and downregulates FASN to promote mitochondrial apoptosis for mammary gland chemoprevention.
Tumor-initiating cells of breast and prostate origin show alterations in the expression of genes related to iron metabolism.
Hydralazine induces stress resistance and extends C. elegans lifespan by activating the NRF2/SKN-1 signalling pathway.
Genetically unmatched human iPSC and ESC exhibit equivalent gene expression and neuronal differentiation potential.
Polyphyllin I Overcomes EMT-Associated Resistance to Erlotinib in Lung Cancer Cells via IL-6/STAT3 Pathway Inhibition.
Compound 49b Restores Retinal Thickness and Reduces Degenerate Capillaries in the Rat Retina following Ischemia/Reperfusion.
Novel biomarkers of resistance of pancreatic cancer cells to oncolytic vesicular stomatitis virus.
Redefining the role of peripheral LPS as a neuroinflammatory agent and evaluating the role of hydrogen sulphide through metformin intervention.
NF-κB and IRF1 Induce Endogenous Retrovirus K Expression via Interferon-Stimulated Response Elements in Its 5' Long Terminal Repeat.
PML plays both inimical and beneficial roles in HSV-1 replication.
An RNA matchmaker protein regulates the activity of the long noncoding RNA HOTAIR.
Effect of β-sitosterol against methyl nitrosourea-induced mammary gland carcinoma in albino rats.
A comparison of genetically matched cell lines reveals the equivalence of human iPSCs and ESCs.
Recombinant H22(scFv) blocks CD64 and prevents the capture of anti-TNF monoclonal antibody. A potential strategy to enhance anti-TNF therapy.
Effects of adult exposure to bisphenol a on genes involved in the physiopathology of rat prefrontal cortex.
Amylin deposition in the brain: A second amyloid in Alzheimer disease?
Impairment of Atg5-dependent autophagic flux promotes paraquat- and MPP⁺-induced apoptosis but not rotenone or 6-hydroxydopamine toxicity.
Gao Y, Wang L, Niu Z, Feng H, Liu J, Sun J, Gao Y, Pan L
Experimental and therapeutic medicine 2022 Feb;23(2):140
Experimental and therapeutic medicine 2022 Feb;23(2):140
Evidence for reprogramming of monocytes into reparative alveolar macrophages in vivo by targeting PDE4b.
Rochford I, Joshi JC, Rayees S, Anwar M, Akhter MZ, Yalagala L, Banerjee S, Mehta D
American journal of physiology. Lung cellular and molecular physiology 2021 Oct 1;321(4):L686-L702
American journal of physiology. Lung cellular and molecular physiology 2021 Oct 1;321(4):L686-L702
Checkpoint inhibition through small molecule-induced internalization of programmed death-ligand 1.
Park JJ, Thi EP, Carpio VH, Bi Y, Cole AG, Dorsey BD, Fan K, Harasym T, Iott CL, Kadhim S, Kim JH, Lee ACH, Nguyen D, Paratala BS, Qiu R, White A, Lakshminarasimhan D, Leo C, Suto RK, Rijnbrand R, Tang S, Sofia MJ, Moore CB
Nature communications 2021 Feb 22;12(1):1222
Nature communications 2021 Feb 22;12(1):1222
Distinct mRNAs in Cancer Extracellular Vesicles Activate Angiogenesis and Alter Transcriptome of Vascular Endothelial Cells.
Zhang P, Lim SB, Jiang K, Chew TW, Low BC, Lim CT
Cancers 2021 Apr 22;13(9)
Cancers 2021 Apr 22;13(9)
Inhalation of lung spheroid cell secretome and exosomes promotes lung repair in pulmonary fibrosis.
Dinh PC, Paudel D, Brochu H, Popowski KD, Gracieux MC, Cores J, Huang K, Hensley MT, Harrell E, Vandergriff AC, George AK, Barrio RT, Hu S, Allen TA, Blackburn K, Caranasos TG, Peng X, Schnabel LV, Adler KB, Lobo LJ, Goshe MB, Cheng K
Nature communications 2020 Feb 28;11(1):1064
Nature communications 2020 Feb 28;11(1):1064
Beneficial Effects of Acetyl-DL-Leucine (ADLL) in a Mouse Model of Sandhoff Disease.
Kaya E, Smith DA, Smith C, Boland B, Strupp M, Platt FM
Journal of clinical medicine 2020 Apr 8;9(4)
Journal of clinical medicine 2020 Apr 8;9(4)
Transcutaneous Vagus Nerve Stimulation Regulates the Cholinergic Anti-inflammatory Pathway to Counteract 1, 2-Dimethylhydrazine Induced Colon Carcinogenesis in Albino wistar Rats.
Rawat JK, Roy S, Singh M, Guatam S, Yadav RK, Ansari MN, Aldossary SA, Saeedan AS, Kaithwas G
Frontiers in pharmacology 2019;10:353
Frontiers in pharmacology 2019;10:353
Hydralazine targets cAMP-dependent protein kinase leading to sirtuin1/5 activation and lifespan extension in C. elegans.
Dehghan E, Goodarzi M, Saremi B, Lin R, Mirzaei H
Nature communications 2019 Oct 28;10(1):4905
Nature communications 2019 Oct 28;10(1):4905
Repurposing mechanistic insight of PDE-5 inhibitor in cancer chemoprevention through mitochondrial-oxidative stress intervention and blockade of DuCLOX signalling.
Singh M, Kasna S, Roy S, Aldosary S, Saeedan AS, Ansari MN, Kaithwas G
BMC cancer 2019 Oct 24;19(1):996
BMC cancer 2019 Oct 24;19(1):996
Activation of prolyl hydroxylase-2 for stabilization of mitochondrial stress along with simultaneous downregulation of HIF-1α/FASN in ER + breast cancer subtype.
Devi U, Singh M, Roy S, Gupta PS, Ansari MN, Saeedan AS, Kaithwas G
Cell biochemistry and function 2019 Jun;37(4):216-227
Cell biochemistry and function 2019 Jun;37(4):216-227
KMT5A promotes metastasis of clear cell renal cell carcinoma through reducing cadherin-1 expression.
Lin ZZ, Ming DS, Chen YB, Zhang JM, Chen HH, Jiang JJ, Zhang ZS
Oncology letters 2019 Jun;17(6):4907-4913
Oncology letters 2019 Jun;17(6):4907-4913
Hyperexcitability of the local cortical circuit in mouse models of tuberous sclerosis complex.
Zhao JP, Yoshii A
Molecular brain 2019 Jan 25;12(1):6
Molecular brain 2019 Jan 25;12(1):6
Developmental NMDA receptor dysregulation in the infantile neuronal ceroid lipofuscinosis mouse model.
Koster KP, Francesconi W, Berton F, Alahmadi S, Srinivas R, Yoshii A
eLife 2019 Apr 4;8
eLife 2019 Apr 4;8
DuCLOX-2/5 Inhibition Attenuates Inflammatory Response and Induces Mitochondrial Apoptosis for Mammary Gland Chemoprevention.
Gautam S, Rawat AK, Sammi SR, Roy S, Singh M, Devi U, Yadav RK, Singh L, Rawat JK, Ansari MN, Saeedan AS, Kumar D, Pandey R, Kaithwas G
Frontiers in pharmacology 2018;9:314
Frontiers in pharmacology 2018;9:314
Ubiquitin-Mimicking Peptides Transfer Differentiates by E1 and E2 Enzymes.
Jin B, Wang J, Liu X, Fang S, Jiang B, Hofmann K, Yin J, Zhao B
BioMed research international 2018;2018:6062520
BioMed research international 2018;2018:6062520
Functional and structural characterization of a novel malignant hyperthermia-susceptible variant of DHPR-β(1a) subunit (CACNB1).
Perez CF, Eltit JM, Lopez JR, Bodnár D, Dulhunty AF, Aditya S, Casarotto MG
American journal of physiology. Cell physiology 2018 Mar 1;314(3):C323-C333
American journal of physiology. Cell physiology 2018 Mar 1;314(3):C323-C333
Selective Cannabinoid 2 Receptor Stimulation Reduces Tubular Epithelial Cell Damage after Renal Ischemia-Reperfusion Injury.
Pressly JD, Mustafa SM, Adibi AH, Alghamdi S, Pandey P, Roy KK, Doerksen RJ, Moore BM Jr, Park F
The Journal of pharmacology and experimental therapeutics 2018 Feb;364(2):287-299
The Journal of pharmacology and experimental therapeutics 2018 Feb;364(2):287-299
Estradiol signaling mediates gender difference in visceral adiposity via autophagy.
Tao Z, Zheng LD, Smith C, Luo J, Robinson A, Almeida FA, Wang Z, Olumi AF, Liu D, Cheng Z
Cell death & disease 2018 Feb 22;9(3):309
Cell death & disease 2018 Feb 22;9(3):309
Evaluation of the effects of the combination of BMP-2-modified BMSCs and PRP on cartilage defects.
Ruan S, Deng J, Yan L, Huang W
Experimental and therapeutic medicine 2018 Dec;16(6):4569-4577
Experimental and therapeutic medicine 2018 Dec;16(6):4569-4577
Disruption of mesoderm formation during cardiac differentiation due to developmental exposure to 13-cis-retinoic acid.
Liu Q, Van Bortle K, Zhang Y, Zhao MT, Zhang JZ, Geller BS, Gruber JJ, Jiang C, Wu JC, Snyder MP
Scientific reports 2018 Aug 28;8(1):12960
Scientific reports 2018 Aug 28;8(1):12960
Human snoRNA-93 is processed into a microRNA-like RNA that promotes breast cancer cell invasion.
Patterson DG, Roberts JT, King VM, Houserova D, Barnhill EC, Crucello A, Polska CJ, Brantley LW, Kaufman GC, Nguyen M, Santana MW, Schiller IA, Spicciani JS, Zapata AK, Miller MM, Sherman TD, Ma R, Zhao H, Arora R, Coley AB, Zeidan MM, Tan M, Xi Y, Borchert GM
NPJ breast cancer 2017;3:25
NPJ breast cancer 2017;3:25
Alpha-linolenic acid stabilizes HIF-1 α and downregulates FASN to promote mitochondrial apoptosis for mammary gland chemoprevention.
Roy S, Rawat AK, Sammi SR, Devi U, Singh M, Gautam S, Yadav RK, Rawat JK, Singh L, Ansari MN, Saeedan AS, Pandey R, Kumar D, Kaithwas G
Oncotarget 2017 Sep 19;8(41):70049-70071
Oncotarget 2017 Sep 19;8(41):70049-70071
Tumor-initiating cells of breast and prostate origin show alterations in the expression of genes related to iron metabolism.
Rychtarcikova Z, Lettlova S, Tomkova V, Korenkova V, Langerova L, Simonova E, Zjablovskaja P, Alberich-Jorda M, Neuzil J, Truksa J
Oncotarget 2017 Jan 24;8(4):6376-6398
Oncotarget 2017 Jan 24;8(4):6376-6398
Hydralazine induces stress resistance and extends C. elegans lifespan by activating the NRF2/SKN-1 signalling pathway.
Dehghan E, Zhang Y, Saremi B, Yadavali S, Hakimi A, Dehghani M, Goodarzi M, Tu X, Robertson S, Lin R, Chudhuri A, Mirzaei H
Nature communications 2017 Dec 20;8(1):2223
Nature communications 2017 Dec 20;8(1):2223
Genetically unmatched human iPSC and ESC exhibit equivalent gene expression and neuronal differentiation potential.
Marei HE, Althani A, Lashen S, Cenciarelli C, Hasan A
Scientific reports 2017 Dec 13;7(1):17504
Scientific reports 2017 Dec 13;7(1):17504
Polyphyllin I Overcomes EMT-Associated Resistance to Erlotinib in Lung Cancer Cells via IL-6/STAT3 Pathway Inhibition.
Lou W, Chen Y, Zhu KY, Deng H, Wu T, Wang J
Biological & pharmaceutical bulletin 2017 Aug 1;40(8):1306-1313
Biological & pharmaceutical bulletin 2017 Aug 1;40(8):1306-1313
Compound 49b Restores Retinal Thickness and Reduces Degenerate Capillaries in the Rat Retina following Ischemia/Reperfusion.
Liu L, Jiang Y, Steinle JJ
PloS one 2016;11(7):e0159532
PloS one 2016;11(7):e0159532
Novel biomarkers of resistance of pancreatic cancer cells to oncolytic vesicular stomatitis virus.
Hastie E, Cataldi M, Moerdyk-Schauwecker MJ, Felt SA, Steuerwald N, Grdzelishvili VZ
Oncotarget 2016 Sep 20;7(38):61601-61618
Oncotarget 2016 Sep 20;7(38):61601-61618
Redefining the role of peripheral LPS as a neuroinflammatory agent and evaluating the role of hydrogen sulphide through metformin intervention.
Tiwari V, Singh M, Rawat JK, Devi U, Yadav RK, Roy S, Gautam S, Saraf SA, Kumar V, Ansari N, Saeedan AS, Kaithwas G
Inflammopharmacology 2016 Oct;24(5):253-264
Inflammopharmacology 2016 Oct;24(5):253-264
NF-κB and IRF1 Induce Endogenous Retrovirus K Expression via Interferon-Stimulated Response Elements in Its 5' Long Terminal Repeat.
Manghera M, Ferguson-Parry J, Lin R, Douville RN
Journal of virology 2016 Oct 15;90(20):9338-49
Journal of virology 2016 Oct 15;90(20):9338-49
PML plays both inimical and beneficial roles in HSV-1 replication.
Xu P, Mallon S, Roizman B
Proceedings of the National Academy of Sciences of the United States of America 2016 May 24;113(21):E3022-8
Proceedings of the National Academy of Sciences of the United States of America 2016 May 24;113(21):E3022-8
An RNA matchmaker protein regulates the activity of the long noncoding RNA HOTAIR.
Meredith EK, Balas MM, Sindy K, Haislop K, Johnson AM
RNA (New York, N.Y.) 2016 Jul;22(7):995-1010
RNA (New York, N.Y.) 2016 Jul;22(7):995-1010
Effect of β-sitosterol against methyl nitrosourea-induced mammary gland carcinoma in albino rats.
Manral C, Roy S, Singh M, Gautam S, Yadav RK, Rawat JK, Devi U, Ansari MN, Saeedan AS, Kaithwas G
BMC complementary and alternative medicine 2016 Jul 29;16:260
BMC complementary and alternative medicine 2016 Jul 29;16:260
A comparison of genetically matched cell lines reveals the equivalence of human iPSCs and ESCs.
Choi J, Lee S, Mallard W, Clement K, Tagliazucchi GM, Lim H, Choi IY, Ferrari F, Tsankov AM, Pop R, Lee G, Rinn JL, Meissner A, Park PJ, Hochedlinger K
Nature biotechnology 2015 Nov;33(11):1173-81
Nature biotechnology 2015 Nov;33(11):1173-81
Recombinant H22(scFv) blocks CD64 and prevents the capture of anti-TNF monoclonal antibody. A potential strategy to enhance anti-TNF therapy.
Hristodorov D, Mladenov R, Brehm H, Fischer R, Barth S, Thepen T
mAbs 2014;6(5):1283-9
mAbs 2014;6(5):1283-9
Effects of adult exposure to bisphenol a on genes involved in the physiopathology of rat prefrontal cortex.
Castro B, Sánchez P, Torres JM, Ortega E
PloS one 2013;8(9):e73584
PloS one 2013;8(9):e73584
Amylin deposition in the brain: A second amyloid in Alzheimer disease?
Jackson K, Barisone GA, Diaz E, Jin LW, DeCarli C, Despa F
Annals of neurology 2013 Oct;74(4):517-26
Annals of neurology 2013 Oct;74(4):517-26
Impairment of Atg5-dependent autophagic flux promotes paraquat- and MPP⁺-induced apoptosis but not rotenone or 6-hydroxydopamine toxicity.
Garcia-Garcia A, Anandhan A, Burns M, Chen H, Zhou Y, Franco R
Toxicological sciences : an official journal of the Society of Toxicology 2013 Nov;136(1):166-82
Toxicological sciences : an official journal of the Society of Toxicology 2013 Nov;136(1):166-82
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Supportive validation
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Beta-Actin was performed by loading 30 µg of various cell lysates per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an HRP-conjugated Beta-Actin monoclonal antibody (Product # MA5-15739-HRP) at a dilution of 1:1000 for 1 hour at room temperature on a rocking platform and washed in TBS-0.1% Tween-20. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).
- Conjugate
- Horseradish Peroxidase
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of A549 (Lane 1), COS-7 (Lane 2), MDCK (Lane 3), C2C12 (Lane 4), MDA-MB-231 (Lane 5), RSC96 (Lane 6) and tissue extracts of Mouse Lung (Lane 7). The blot was probed with beta Actin Monoclonal Antibody (BA3R), HRP (Product # MA5-15739-HRP, 1 µg/mL) and detected by chemiluminescence. A 42 kDa band corresponding to beta Actin was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
- Conjugate
- Horseradish Peroxidase
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
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- Horseradish Peroxidase
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- Invitrogen Antibodies (provider)
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- Horseradish Peroxidase
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- Horseradish Peroxidase
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- Invitrogen Antibodies (provider)
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- Horseradish Peroxidase
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- Invitrogen Antibodies (provider)
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- Horseradish Peroxidase
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- Invitrogen Antibodies (provider)
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- Horseradish Peroxidase
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- Invitrogen Antibodies (provider)
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- Horseradish Peroxidase
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- Invitrogen Antibodies (provider)
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- Horseradish Peroxidase
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- Horseradish Peroxidase
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- Invitrogen Antibodies (provider)
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- Horseradish Peroxidase
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- Invitrogen Antibodies (provider)
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- Conjugate
- Horseradish Peroxidase
- Submitted by
- Invitrogen Antibodies (provider)
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- Figure 11 ALA mediated activation of mitochondrial associated protein signaling in mammary gland cells Protein extracted from individual groups [1-control, 2-DMBA treated, 3-ALA (0.25 ml/kg, p.o. + DMBA 8 mg/kg, i.v.) and 4- ALA (0.5 ml/kg, p.o. + DMBA 8 mg/kg, i.v.)] were subjected to immunoblotting of proapoptotic (BAX) and anti-apoptotic (Bcl-2 and Bcl-xl) protein with downstream apoptotic markers (VDAC, cytochrome-c, Apaf-1and procaspase9) of respective pathway. mRNA expression of above mentioned protein were also in line with the findings of immunoblotting assay. beta-actin was used as loading control. Each experiment was performed in triplicate. Values are presented as mean +- SD. Comparisons are made by the one-way ANOVA followed by Bonferroni multiple test. All groups are compared to the DMBA treated group (*p
- Conjugate
- Horseradish Peroxidase
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- Fig. 4 Western Blot analysis: Group I: Control; Group II: MNU; Group III: beta-sitosterol10 mg/kg; Group IV: beta-sitosterol 20 mg/kg. The western blot analysis containing whole tissue lysate from rat mammary gland was probed for P-gp 9.5 antibody; its immunoreactivity was evident as a band of molecular weight of 27 kDa. In case of control the expression is low in comparison with the toxicant. The low and high dose of beta-sitosterol shows a similar pattern of elevated expression in a dose dependent manner. Results of the beta-actin analysis are shown as an internal control
- Conjugate
- Horseradish Peroxidase
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 12 Effect of ALA on HIF-1alpha mediated hypoxic markers and fatty acid synthesis regulators Immunoblotting of respective individual group [1-control, 2-DMBA treated, 3-ALA (0.25 ml/kg, p.o. + DMBA 8 mg/kg, i.v.) and 4- ALA (0.5 ml/kg, p.o. + DMBA 8 mg/kg, i.v.)] for HIF-1alpha, PHD2, and FASN conclude the hypoxic microenvironment after DMBA treatment. Perspective correlations of NFkappaBp65, UCHL-1 and SREBP-1c with the above mentioned protein are well defined through the experimental pathway. Excised mammary gland tissue sample lysed in trizol for RNA extraction and analyzed for the mRNA expression of HIF-1alpha, PHD2, FASN, NFkappaBp65, UCHL-1 and SREBP-1c by qRT-PCR: fold induction is relative to tissue under hypoxic conditions after normalization to the beta-actin expression. Each experiment was performed in triplicate. Values are presented as mean +- SD. Comparisons are made on the basis of the one-way ANOVA followed by Bonferroni multiple test. All groups are compared to the DMBA treated group (*p
- Conjugate
- Horseradish Peroxidase
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- 4 BBAP-2-mediated activation of mitochondrial associated protein signalling in mammary gland cells. Protein extracted from individual groups (group 1--control [0.9% normal saline, p.o. ]; group 2--toxic control [MNU, 8 mg/kg, i.v. ]; group 3--BBAP-2 + MNU [68.52 mug/kg, s.c. + 8 mg/kg MNU, i.v. ]; and group 4--BBAP-2 + MNU [137.04 mug/kg, s.c. + 8 mg/kg MNU, i.v.]) was subjected to immunoblotting of antiapoptotic (Bcl-2 and Bcl-xl) and proapoptotic (BAX) proteins with downstream apoptotic markers (VDAC, cytochrome-c, Apaf-1, and procaspase 9) of mitochondrial mediated apoptotic pathway. mRNA expression of above-mentioned protein was also done by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). beta-actin was used as internal loading control. Treatment with BBAP-2 decreased the expression of antiapoptotic protein Bcl-xl without imparting any significant effect upon Bcl-2 and BAX (proapoptotic) while induction (BAX) and inhibition (Bcl-xl and Bcl-2) were observed when quantified through qRT-PCR assay. Each experiment was performed in triplicate. Values are presented as mean +- SD. Each group contains eight animals. Comparisons are made on the basis of the one-way ANOVA followed by Bonferroni multiple test. All groups are compared with the toxic control group (* p< 0.05, ** p< 0.01, *** p< 0.001)
- Conjugate
- Horseradish Peroxidase
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- 5 Downregulation of hypoxic pathway in mammary gland cancer. Immunoblotting of respective individual group (group 1--control [0.9% normal saline, p.o. ]; group 2--toxic control [MNU, 8 mg/kg, i.v. ]; group 3--BBAP-2 + MNU [68.52 mug/kg, s.c. + 8 mg/kg MNU, i.v. ]; and group 4--BBAP-2 + MNU [137.04 mug/kg, s.c. + 8 mg/kg MNU, i.v.]) was performed for estimation of HIF-1alpha, PHD2, and FASN as well as PHD2 in tumour growth. Perspective correlations of NFkappaBp65, UCHL-1, and SREBP-1c with the above-mentioned protein was well defined through the experimental pathway. Excised mammary gland tissue sample lyzed in trizol for RNA extraction and analysed for the mRNA expression of HIF-1alpha, PHD2, FASN, NFkappaBp65, UCHL-1, and SREBP-1c by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR): fold induction is relative to tissue under hypoxic conditions after normalization to the beta-actin expression. Columns: mean of three experimental determination; bars, SD. Comparisons are made on the basis of the one-way ANOVA followed by Bonferroni multiple test. All groups are compared with the toxic control group (* p< 0.05, ** p< 0.01, *** p< 0.001)
- Conjugate
- Horseradish Peroxidase
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- Figure 2 Effect of acetyl- DL -leucine on autophagy in the cerebellum of 12-week-old Hexb -/- mice. Wild-type untreated ( Hexb +/+ ), Sandhoff untreated ( Hexb -/- UT), ADLL ( Hexb -/- ADLL). In total, four animals for Hexb +/+ , and five animals for Hexb -/- UT and Hexb -/- ADLL. Proteins of interests were probed by Western blot and normalized to beta actin. Fluorescence values were obtained with Fiji software and calculated relative to Hexb +/+ . ( a ) mTOR, p-mTOR expressions and mTOR p :T rations, mean +- SD * p < 0.05, (two-way ANOVA). ( b ) Western blot images of p -mTOR and mTOR proteins and their beta-actin loading controls. ( c ) LC3-I, LC3-II expressions, and ratio of LC3-II versus LC3-I, mean +- SD, * p = 0.0483, ** p = 0.0011, *** p = 0.0006, **** p < 0.0001 (two-way ANOVA). ( d ) p62 levels, mean +- SD (one-way ANOVA). ( e ) Western blot images of LC3s and p62 with their beta-actin loading controls. Samples on the same blots were cut (inserted lines) for appropriate visual representation.
- Conjugate
- Horseradish Peroxidase
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- Invitrogen Antibodies (provider)
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- Figure 3 Effects of acetyl-DL-leucine on BCAA and glutamate and energy metabolism in the cerebellum of 12-week-old Hexb -/- mice. ( a ) BCKADH, p -BCKADH, p /t BCKADH expressions, mean +- SD, * p = 0.0494 (two-way ANOVA). ( b ) GDH expression. Mean +- SD ** p < 0.0057 (one-way ANOVA). ( c ) PGC1-alpha expression, mean +- SD (one-way ANOVA) ( d ) BCKADH, p-PCKADH, GDH and PGC1-alpha Western blot images with beta-actin loading controls. Samples on the same blots were cut (inserted lines) for appropriate visual representation.
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- Figure 4 Effects of acetyl-DL-leucine on glucose and antioxidant metabolism in the cerebellum of 12-week-old Hexb -/- mice. ( a ) PDHC protein expression, mean +- SD, * p = 0.0105, ** p = 0.0097, *** p = 0.001 (two-way ANOVA). ( b ) p -PDH expression, mean +- SD, * p = 0.0310, ** p = 0.0062 (two-way ANOVA). ( c ) p-PDH/-tPDHE1alpha ratios, mean +- SD, * p < 0.023 (two-way ANOVA). ( d ) LDH expression, mean +- SD, ** p = 0.0071 (one-way ANOVA). ( e ) PDK 1-2-4 and PDP1 expression, mean +- SD, *** p = 0.0002, **** p < 0.0001 (two-way ANOVA). ( f ) Western blot images of PDHC, p-PDH, LDH, PDKs and PDP1 along with their beta-actin controls. ( g ) SOD1 and SOD2 expression, mean +- SD, * p = 0.0216, ** p = 0.0121, **** p < 0.0001 (two-way ANOVA). ( h ) Western blot images of SOD1 and SOD2 and their beta-actin loading controls. Samples on the same blots were cut (inserted lines) for appropriate visual representation.
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- FIGURE 3 Effect of taVNS on mitochondrial apoptosis. The details of the groups for western blot analysis and caspase assay are as follows: I-Control, II-taVNS control, III-DMH control, IV-taVNS1, V-taVNS 2, VI-taVNS 3, VII-taVNS 4, VIII-taVNS 5, IX-Standard chemotherapy, X-Dummy control. Specific treatments for particular groups mentioned in Supplementary Table 1 . (A) Proteins were extracted from individual groups and subjected to immunoblotting of pro-apoptotic (BAX and BAD) and anti-apoptotic (Bcl-2 and Bcl-xl) markers along with downstream markers of apoptotic pathway (VDAC, cytochrome-c, Apaf-1, and procaspase 9). After treatment with DMH the anti-apoptotic proteins (Bcl-2 and Bcl-xl) expression was increased. The expression of pro-apoptotic protein BAX decreased while BAD increased. The taVNS treatment results in restoration of anti-apoptotic and pro-apoptotic proteins with exception of BAD. When observed, the expression of markers of mitochondrial apoptosis (VDAC, cytochrome c, Apaf-1, and procaspase 9) treatment with DMH results in increased expression of VDAC, Apaf-1, and procaspase 9 along with decreased expression of cytochrome c. Treatment with taVNS resulted in increased expression of cytochrome c and decreased expression of procaspase 9. (B) The outcomes from the immunoblotting assay were confirmed through qRT-PCR studies by scrutinizing the respective phenotypes of pathway associated proteins. The mRNA expressions of the above mentioned protein were also in l
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- Figure 5 Downstream effects of cancer EVs on angiogenesis. ( A ) Tube formation assay of HUVECs incubated in basal medium (control), treated with 50 ug/mL MDA231-HEVs, or treated with both MDA231-HEVs and 0.1 uM Axitinib (inhibitor of VEGFR) for 24 h. The EVs induced formation of denser tube networks, which can be efficaciously blocked by Axitinib. Images are representative of two independent experiments. ( B ) The total tube length of images from the same experiment were analyzed by ImageJ and normalized by the average of the control group ( N = 6). ( C ) MDA231-HEVs upregulated the VEGF-VEGFR angiogenesis pathway related gene, while showing no effect on the other ANGPT-TIE angiogenesis pathway in HUVECs ( N = 3). ( D ) Western blot of proteins involved in the VEGF-VEGFR angiogenesis pathway. MDA231-HEVs conspicuously enhanced the level of VEGFR1 and the phosphorylation of downstream Erk1/2. Average intensities of the bands were first normalized to beta-actin, and then normalized to the control group. Images are representative of two independent experiments. n.s. : not significant, * P < 0.05, ** P < 0.01 based on Mann-Whitney U test for ( B ), or Student's t test for ( C ). Error bars stand for SEM for ( B , C ).
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- Figure 8. Lentivirus-mediated expression of BMP-2 in bone marrow-derived stromal cells is observed for at least 16 weeks. (A) Protein was isolated from articular cartilage and subchondral bone areas from each group in week 16. Western blot analysis was performed to detect the protein expression of BMP-2 and SOX9. (B) Relative expression of proteins normalized by beta-actin (n=3; *P
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- Fig. 8 Tadalafil mediated activation of mitochondrial mediated apoptosis pathway. Protein extracted from individual groups [Control (Normal saline, 3 ml/kg, p.o.); Toxic control (MNU 47 mg/kg, i.v.); MNU + Tadalafil (47 mg/kg i.v. + 2 mg/kg p.o.) and MNU + Tadalafil (47 mg/kg i.v. + 4 mg/kg p.o.)] were subjected to immunoblotting of proapoptotic (BAD) and anti-apoptotic (Bcl-xl) marker along with COX, NFkappaBp65 and UCHL-1. beta-actin was used as internal loading control. Each experiment was performed in triplicate. The data was represented as mean +- SD. The groups were significantly different by one-way ANOVA followed by Bonferroni multiple tests. All groups were compared to the toxic control group (* p < 0.05, ** p < 0.01, *** p < 0.001)
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- Fig. 2 Hydralazine activates SIRT1 and SIRT5 to promote mitochondrial function. a Hydralazine treatment (10 uM) increases the expression of SIRT5 mRNA in SH-SY5Y cells measured by qPCR using actin as an internal control ( n = 3, mean +- SEM). b Hydralazine increases protein abundance of SIRT1, SIRT5, and TFAM demonstrated by Western blot analysis. Cells were treated with different doses of hydralazine or 20 uM of resveratrol or isoniazid as positive and negative controls for 24 h ( n = 3, mean +- SD). c SIRT1 enzymatic activity was measured by monitoring the fluorescence emission of an acetylated peptide substrate in C2C12 cells treated with hydralazine for 48 h ( n = 4, mean +- SD). d PGC1A immunoprecipitation followed by Western blot analysis showing activation of PGC1A in hydralazine treated SH-SY5Y cells ( n = 3, mean +- SD). e Western blot analysis demonstrates a SIRT5-dependent increase in deacetylation of CPS1 with hydralazine treatment. 5 uM resveratrol used as a positive control ( n = 2-3, mean +- SD). f Flow cytometric quantification of DeltaPsim indicates activation of mitochondria in a SIRT1 and SIRT5-dependent manner ( n = 2, mean +- SD). g Seahorse XF assay demonstrates a SIRT1 and SIRT5-dependent increase in oxygen consumption rate in SH-SY5 cells treated with hydralazine (5 uM, 3 days) ( n = 3, mean +- SEM). h Western blot analysis demonstrating the effects of hydralazine-induced SIRT1 activation on the abundances of SIRT5 and mitochondrial markers in C2C12 ce
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- Figure 11. Hyperpalmitoylation of Fyn kinase and GluN2B is reversed in Ppt1 -/- primary cortical neurons by palmitoylation inhibitor treatment. ( A ) Representative post-APEGS immunoblot of PSD-95 with beta-actin loading control. ( B ) Quantification of total PSD-95 levels following chronic (7d) treatment with vehicle or the palmitoylation inhibitors, 2 BP (1 muM) or cerulenin (1 muM) where indicated. ( C ) Quantification of the ratio of palmitoylated/non-palmitoylated PSD-95 levels following chronic (7d) treatment with vehicle or the palmitoylation inhibitors, 2 BP (1 muM) or cerulenin (1 muM) where indicated. ( D ) Representative post-APEGS immunoblot of Fyn kinase with beta-actin loading control and minus hydroxylamine (-HA) control. ( E ) Quantification of total Fyn kinase levels following chronic (7d) treatment with vehicle or the palmitoylation inhibitors, 2 BP (1 muM) or cerulenin (1 muM) where indicated. ( F ) Quantification of the ratio of palmitoylated/non-palmitoylated Fyn kinase levels following chronic (7 days) treatment with vehicle or the palmitoylation inhibitors, 2 BP (1 muM) or cerulenin (1 muM) where indicated. ( G ) Representative post-APEGS immunoblot of GluN2B with beta-actin loading control and minus hydroxylamine (-HA) control. ( H ) Quantification of total GluN2B levels following chronic (7d) treatment with vehicle or the palmitoylation inhibitors, 2 BP (1 muM) or cerulenin (1 muM) where indicated. ( I ) Quantification of the ratio of palmitoylated
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- Figure 2--figure supplement 1. NMDAR subunit composition is immature in whole lysates from Ppt1 -/- visual cortex. ( A ) Representative immunoblots of the GluN2A in whole lysates across age and genotype as indicated (top) and quantification of band density (bottom) normalized to beta-actin loading control within lane. ( B ) Representative immunoblots of the GluN2B in whole lysates across age and genotype as indicated (top) and quantification of band density (bottom) normalized to beta-actin loading control within lane. ( C ) Representative immunoblots of GluN2A and GluN2B (top) from whole lysates across age and genotype and quantification of the ratio of GluN2A/GluN2B band density within animal (bottom). ( D ) Representative immunoblots of GluN1 in whole lysates across age and genotype as indicated (top) and quantification of band density (bottom) normalized to beta-actin loading control within lane. ( E ) Representative immunoblot from whole lysates of PPT1 across age and genotype as indicated (top) and protein expression level (bottom) normalized to beta-actin. For experiments in Figure 2--figure supplement 1A-C , Ppt1 -/- and WT were compared (n = 4 independent experiments/animals with two repetitions/group) at each age using t-test and the significance is indicated: *p
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- Fig. 2 LSC-Sec inhalation treatment promotes alveolar repair. a Representative immunostaining of lung sections for von Willebrand Factor (vWF), pro-surfactant protein C (Pro-SPC) and aquaporin 5 (AQP5). Scale bar = 100 mum. b - d Quantification of percent pixel intensity of vWF+ ( b ), percent ProSPC+ nuclei ( c ), and percent pixel intensity of AQP5+ ( d ); each dot represents data from one animal; n = 12 biological independent animals. e - f Immunoblot analysis of aquaporin 5 (AQP5), pro-surfactant protein C (Pro-SPC), von Willebrand Factor (vWF), alpha smooth muscle actin (alphaSMA), SMAD3, matrix metalloproteinase 2 (MMP-2), and beta-actin loading control (B-actin) from lung protein lysate ( e ) with corresponding quantification of protein levels as fold of sham control ( f ); each dot represents data from one animal; n = 3 biological independent animals. g - h Representative cytokine array with quantification of relative intensity ( g ) and corresponding quantification of relative intensity ( h ). Throughout, data are mean +- s.d. P -value as indicated by non-parametric one-way ANOVA.
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- Figure 6 Expression level of protein of COX-2 and 5-LOX through western blot and levels of gene contributor through quantitative RT-PCR. Immunoblotting of respective individual group [I-Control (Normal saline, 3 ml/kg, p.o.), II- Toxic control (MNU 47 mg/kg, i.v.), III- Zaltoprofen (10 mg/kg, p.o. + MNU 47 mg/kg, i.v.), IV- Zileuton (10 mg/kg, p.o. + MNU 47 mg/kg, i.v.) and V- Zaltoprofen + Zileuton (5 + 5 mg/kg, p.o. + MNU 47 mg/kg, i.v.)] for COX-2 and 5-LOX. Excised mammary gland tissue sample lysed in trizol for RNA extraction and analyzed for the mRNA expression of COX-2 and 5-LOX by qRT-PCR. beta-actin was used as loading control. Each experiment was performed in triplicate. Values are presented as Mean +- SD. Comparisons were made by the one-way ANOVA followed by Bonferroni multiple test. All groups were compared to the MNU treated group ( ** p < 0.01, *** p < 0.001).
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