Antibody data
- Antibody Data
- Antigen structure
- References [6]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
- Flow cytometry [1]
- Other assay [3]
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Validation data
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- Product number
- 44-1150G - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-AMPK alpha-1,2 (Thr183, Thr172) Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Storage
- -20°C
Submitted references Glycocalyx heparan sulfate cleavage promotes endothelial cell angiopoietin-2 expression by impairing shear stress-related AMPK/FoxO1 signaling.
ACSL4 contributes to sevoflurane-induced ferroptotic neuronal death in SH-SY5Y cells via the 5' AMP-activated protein kinase/mammalian target of rapamycin pathway.
3H-1,2-Dithiole-3-Thione Protects Lens Epithelial Cells against Fructose-Induced Epithelial-Mesenchymal Transition via Activation of AMPK to Eliminate AKR1B1-Induced Oxidative Stress in Diabetes Mellitus.
AKR1B1-Induced Epithelial-Mesenchymal Transition Mediated by RAGE-Oxidative Stress in Diabetic Cataract Lens.
N-myristoyltransferase deficiency impairs activation of kinase AMPK and promotes synovial tissue inflammation.
Impaired Cellular Energy Metabolism Contributes to Duck-Enteritis-Virus-Induced Autophagy via the AMPK-TSC2-MTOR Signaling Pathway.
Richter RP, Ashtekar AR, Zheng L, Pretorius D, Kaushlendra T, Sanderson RD, Gaggar A, Richter JR
JCI insight 2022 Aug 8;7(15)
JCI insight 2022 Aug 8;7(15)
ACSL4 contributes to sevoflurane-induced ferroptotic neuronal death in SH-SY5Y cells via the 5' AMP-activated protein kinase/mammalian target of rapamycin pathway.
Cheng L, Zhu X, Liu Y, Zhu K, Lin K, Li F
Annals of translational medicine 2021 Sep;9(18):1454
Annals of translational medicine 2021 Sep;9(18):1454
3H-1,2-Dithiole-3-Thione Protects Lens Epithelial Cells against Fructose-Induced Epithelial-Mesenchymal Transition via Activation of AMPK to Eliminate AKR1B1-Induced Oxidative Stress in Diabetes Mellitus.
Wu TT, Chen YY, Ho CY, Yeh TC, Sun GC, Tseng CJ, Cheng PW
Antioxidants (Basel, Switzerland) 2021 Jul 6;10(7)
Antioxidants (Basel, Switzerland) 2021 Jul 6;10(7)
AKR1B1-Induced Epithelial-Mesenchymal Transition Mediated by RAGE-Oxidative Stress in Diabetic Cataract Lens.
Wu TT, Chen YY, Chang HY, Kung YH, Tseng CJ, Cheng PW
Antioxidants (Basel, Switzerland) 2020 Mar 25;9(4)
Antioxidants (Basel, Switzerland) 2020 Mar 25;9(4)
N-myristoyltransferase deficiency impairs activation of kinase AMPK and promotes synovial tissue inflammation.
Wen Z, Jin K, Shen Y, Yang Z, Li Y, Wu B, Tian L, Shoor S, Roche NE, Goronzy JJ, Weyand CM
Nature immunology 2019 Mar;20(3):313-325
Nature immunology 2019 Mar;20(3):313-325
Impaired Cellular Energy Metabolism Contributes to Duck-Enteritis-Virus-Induced Autophagy via the AMPK-TSC2-MTOR Signaling Pathway.
Yin H, Zhao L, Li S, Xu L, Wang Y, Chen H
Frontiers in cellular and infection microbiology 2017;7:423
Frontiers in cellular and infection microbiology 2017;7:423
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Upregulation, Antibody-Peptide Competition and Phosphatase Stripping. Extracts of HepG2 cells untreated (1) or treated with 12 mM Metformin for 24 hours in serum free media (2-6) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was left untreated (1-5) or treated with lambda phosphatase (6), blocked with a 3% BSA-TBST buffer for one hour at room temperature, and then incubated with the Phospho-AMPK alpha-1,2 (Thr183, Thr172) antibody (Product # 44-1150G) for two hours at room temperature in 3% BSA-TBST buffer, following prior incubation with: no peptide (1,2), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), a generic phosphothreonine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and signals were detected using the Pierce SuperSignal™ method. The data show that only the phosphopeptide corresponding to Phospho-AMPK alpha-1,2 (Thr183, Thr172) completely blocks the signal and that phosphatase stripping eliminates the signal, verifying that the antibody is phosphorylation site-specific. The data also show upregulation of the phospho-signal upon Metformin treatment in this cell system.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Phospho-AMPK alpha 1,2 (Thr183, Thr172) was done on 70% confluent log phase MDA-MB-231 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Phospho-AMPK alpha-1,2 (Thr183, Thr172) Rabbit Polyclonal Antibody (Product # 44-1150G) at 1:250 dilution in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing Nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of Phospho-AMPK alpha 1,2 (Thr183, Thr172) showing staining in the cytoplasm of paraffin-embedded human brain tissue (right) compared to 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 Phospho-AMPK alpha-1,2 (Thr183, Thr172) Rabbit Polyclonal Antibody (Product # 44-1150G) diluted in 3% BSA-PBS at a dilution of 1:20 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 AMPK alpha 1 + 2 [pThr172] was done on MDA-MB-231 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with AMPK alpha 1 + 2 [pThr172] Rabbit Polyclonal Antibody (441150G, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 4 D3T reduces fructose-induced EMT progression in the lens of rats with fructose-induced type 2 DM. ( A ) Immunoblotting analysis depicting Aldose reductase, p-AMPK T172 and Ac-SOD2 expression in the fructose-induced type 2 DM lens with or without D3T administration. ( B ) Immunoblotting analysis showing Snail and Slug protein expressions in the fruc Table 2 . DM lens with or without D3T administration. ( C ) Protein expressions of E-cadherin and vimentin were also analyzed and quantified. All data represented as mean +- SEM ( n = 6 per group, independent experiments in each figure).