MA5-15739

antibody from Invitrogen Antibodies

Targeting: ACTB

Provider product page for MA5-15739

Western blot

ELISA

Antibody data

Antibody Data
Antigen structure
References [312]
Comments [0]
Validations
Western blot [5]
Immunocytochemistry [1]
Immunohistochemistry [4]
Flow cytometry [3]
Other assay [25]

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Product number: MA5-15739 - Provider product page
Provider: Invitrogen Antibodies
Product name: beta Actin Loading Control Monoclonal Antibody (BA3R)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: Storage and handling: add 0.05% sodium azide if desired. For long term storage, store at -20ºC, avoiding freeze/thaw cycles.
Reactivity: Human, Mouse, Rat, Chicken/Avian, Rabbit
Host: Mouse
Isotype: IgG
Antibody clone number: BA3R
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

ACTB protein structure - MA5-15739 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 1:1000 (1 µg/mL) Ab dilution used in WB of 20 µg/lane tissue lysates from human (1), mouse (2), rat (3), and rabbit (4)

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Beta-Actin was performed by loading 20 µg of THP-1 whole cell lysate per well onto a SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 1 hour at room temperature. The membrane was probed with a Beta-Actin monoclonal antibody (Product # MA5-15739) at a dilution of 1:5000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated goat anti-mouse IgG secondary at a dilution of 1:40,000 for 1 hour at room temperature. Chemiluminescent detection was performed using ECL substrate. Data courtesy of the Innovators Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of beta-actin was performed by loading 8 µg of mouse embryonic stem cell lysate per well onto an SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% milk in TBST for one hour at room temperature. The membrane was probed with a beta-actin loading control monoclonal antibody (Product # MA5-15739) at a dilution of 1:5000 for two hours at room temperature, washed in TBST, and probed with an HRP-conjugated anti-mouse IgG secondary antibody at a dilution of 1:25,000 for one hour at room temperature. Detection was performed using a chemiluminescent substrate. Data courtesy of the Innovators Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 1:1000 (1 µg/mL) Ab dilution used in WB of 20 µg/lane tissue lysates from human (1), mouse (2), rat (3), and rabbit (4)

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30µg lysate) of A-431 (Lane 1), COS-7 (Lane 2), MDCK (Lane 3), C2C12 (Lane 4), PC-3 (Lane 5), L6 (Lane 6), RSC96 (Lane 7), tissue extracts (30µg lysate) of Mouse Lung (Lane 8), Rat Stomach (Lane 9), Mouse Pancreas (Lane 10) and Rat Brain (Lane 11). The blots were probed with Anti-beta Actin Mouse Monoclonal Antibody (Product # MA5-15739-1MG, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/mL, 1:4000 dilution). 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® 4-12 % Bis-Tris gel (Product # NP0321BOX), 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 and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Beta Actin showing staining in the cytoskeleton of paraffin-embedded human breast carcinoma (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Beta Actin loading control antibody (Product # MA5-15739) diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Beta Actin showing staining in the cytoskeleton of paraffin-embedded human skeletal muscle tissue (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Beta Actin loading control antibody (Product # MA5-15739) diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Beta Actin showing staining in the cytoskeleton of paraffin-embedded mouse skeletal muscle tissue (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Beta Actin loading control antibody (Product # MA5-15739) diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Beta Actin showing staining in the cytoskeleton of paraffin-embedded mouse skeletal muscle tissue (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Beta Actin loading control antibody (Product # MA5-15739) diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Beta Actin in A431 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a Beta Actin loading control antibody (Product # MA5-15739) at a dilution of 2 µg/test for 40 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Beta Actin in Hela cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a Beta Actin loading control antibody (Product # MA5-15739) at a dilution of 2 µg/test for 40 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Beta Actin in NIH-3T3 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a Beta Actin loading control antibody (Product # MA5-15739) at a dilution of 2 µg/test for 40 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 p21 degradation is mediated by the CRL4 Cdt2 ligase complex. A ) Schematic illustrating the experimental protocol for siRNA knockdown of ligase components in Figures 1B and 1C . B and C ) p21 degradation requires DDB1 (B) and Cdt2 (C) . Murine A9 cells were targeted with control siRNA or siRNA to DDB1 (B) or Cdt2 (C) as depicted in Figure 1A . Uninfected control cells were harvested at the time of release (Mock T0). Infections were done at the time of release at an MOI of 10 before harvest at 24 and 40 hr pi. Western blots were performed using antibodies against the indicated proteins.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 The CRL4 Cdt2 E3 ligase complex is important for MVM replication. A ) Schematic showing the experimental protocol for figures 2B and 2C . B and C ) Upper panels: murine A9 cells treated with siRNA as shown in 2A were infected at an MOI of 0.5, harvested at the indicated time points and processed for Southern blotting using an MVM genomic probe. Rf - replicative forms. SS - single stranded genomic DNA. Representative Southern Blots are shown; quantifications in the text reflect two DDB1 and three Cdt2 separate knockdown experiments. Lower panels: western blots show knockdown of DDB1 and Cdt2 done in parallel experiments under identical conditions to replication assays.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2. AS1842856 suppressed PPARgamma and mitochondrial protein expression. ( A ) Western blots showing the effect of AS1842856 on PPARgamma, adiponectin, mitochondrial proteins C1 and C3. beta-actin was probed as the loading control. DI, differentiation induction; AS, AS1842856. ( B-D ) Densitometric analysis of western blot images with NIH ImageJ software; n = 3-5. * P < 0 .05; **, P < 0 .01; and ***, P < 0 .0001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4. Kinetics of FoxO1-regulated protein expression during adipogenesis. ( A ) Western blots showing the expression of PPARgamma, adiponectin, mitochondrial proteins C1 and C3. beta-actin was probed as the loading control. ( B ) Densitometric analysis of western blot images for PPARgamma and adiponectin with NIH ImageJ software. ( C ) Densitometric analysis of protein gel blot images for C1 and C3 with NIH ImageJ software. n = 3-5. * P < 0 .05; NS, not significant.

Supportive validation

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Experimental details: Fig. 4 Effects of BME on hepatic metabolic signaling pathway. (A) protein expressions of liver SIRT1 (B), phosphorylated AMPK, ACC (C),SIRT1, Foxo1, PPARalpha and SREBP1c were determined by Western blotting. HFDwas used as a control and beta-actin, alpha- tubulin ware used as a loadingcontrol.

Supportive validation

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Experimental details: 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

Supportive validation

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Experimental details: Figure 3 Differentially expressed genes (DEGs) between isogenic hESC and hiPSC lines (a) Venn diagram showing the number of genes consistently up- or down- regulated in 3 biological replicate hiPSC lines when compared to 3 biological replicate hESC GFP lines from the same genetic background. (FDR

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 5 Western blot analysis fold expression of HSP90 calculated using the beta-actin as an internal standard. a) HEK-293 cells treated with C - Control (0 mug/mL), 40,80,160 and 320 mug/mL lemongrass oil b) MCF-7 cells treated with C - Control (0 mug/mL), 40, 80, 160 and 320 mug/mL lemongrass oil. Fig. 5

Supportive validation

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Experimental details: 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