Anti-AMPK alpha 1 antibody

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMPK alpha 1 antibody (AB3759)

IHC image of AMPK alpha 1 staining in Human normal pancreas formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab3759, 5μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMPK alpha 1 antibody (AB3759)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human muscle tissue labelling AMPK alpha 1 with ab3759 at 1/5000 (0.2µg/ml). Detection : DAB.

• IP

Supplier Data

Immunoprecipitation - Anti-AMPK alpha 1 antibody (AB3759)

Samples : Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer.

Lane 1 and 2 : ab3759 used for IP at 3 μg per reaction, two lots.

Lane 3 : Control IgG.

For blotting immunoprecipitated AMPK alpha 1, ab3759 was used at 0.4 μg/ml.

Detection : Chemiluminescence with an exposure time of 3 seconds.

All lanes:

Immunoprecipitation - Anti-AMPK alpha 1 antibody (ab3759)

Predicted band size: 64 kDa

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• WB

Supplier Data

Western blot - Anti-AMPK alpha 1 antibody (AB3759)

Lysated prepared using NETN lysis buffer.

All lanes:

Western blot - Anti-AMPK alpha 1 antibody (ab3759) at 0.1 µg/mL

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 15 µg

Lane 2:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 15 µg

Lane 3:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 15 µg

Lane 4:

TCMK-1 (Mouse kidney epithelial cell line) whole cell lysate at 15 µg

Lane 5:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 15 µg

Predicted band size: 64 kDa

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Exposure time: 3s

• WB

Unknown

Western blot - Anti-AMPK alpha 1 antibody (AB3759)

Samples : Extracts from

1. Bovine aortic endothelial cells

2. Rat aortic smooth muscle cells

3. HepG2 cells

4. Human aortic endothelial cells

Antibody : Affinity purified Rabbit anti-AMPK alpha 1 (ab3759) used at 2ug/ml (1/500).

All lanes:

Western blot - Anti-AMPK alpha 1 antibody (ab3759)

Predicted band size: 64 kDa

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• WB

CiteAb

Western blot - Anti-AMPK alpha 1 antibody (AB3759)

AMPK alpha 1 western blot using anti-AMPK alpha 1 antibody ab3759. Publication image and figure legend from Wu, L., Zhang, L., et al., 2018, Front Physiol, PubMed 29515462.

ab3759 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab3759 please see the product overview.

Adipose AMPK deficiency impaired cold tolerance and suppressed cold-induced thermogenesis specifically in inguinal white adipose tissue. (A) Western blot analysis of UCP1, p-AMPKα (T172) and AMPKα protein levels in the interscapular brown adipose tissue (BAT), inguinal white adipose tissue (iWAT) and epididymal white adipose tissue (eWAT) of 9-week-old male wild-type (WT) mice housed at RT or 4°C for 24 h. β-actin was used as a loading control. n = 4. (B) Western blot analysis of AMPKα1, α2, β1, β2, γ1, γ2 protein expression levels in the BAT, iWAT, eWAT and liver of 10-week-old male AKO mice and age-matched floxed littermates. β-actin was used as a loading control. n = 3. (C) Rectal temperature of 8-week-old chow-fed AKO and floxed mice at 4°C for 8 h. n = 9. (D–M) 8-week-old male chow-fed AKO mice and floxed mice were housed at 4°C for 48 h. Representative transmission electronic microscopy images of iWAT (D) and BAT (G), total number of mitochondria per micrograph in iWAT (E) and BAT (H) and the percentage of mitochondria with disrupted cristae over total mitochondria in iWAT (F) and BAT (I) from AKO mice and floxed mice were shown. n = 3. Scale bar : 2 μm in low magnification (×5,000, upper) and 0.5 μm in high magnification (×20,000, bottom). The relative mRNA levels of thermogenic genes and fatty acid oxidation (FAO)-related genes in the iWAT (J) and BAT (K) of AKO mice and floxed mice were analyzed by quantitative RT-PCR (normalized to 36b4). n = 8–9. The expression levels of AMPKα, p-AMPKα (T172), ACC, p-ACC (S79), UCP1 and PGC-1α in the iWAT (L) and BAT (M) of AKO mice and floxed mice were determined by western blot analysis. Data are presented as the means ± SEM. Student's t-test. *p < 0.05, **p < 0.01, ***p < 0.001 compared with the indicated control group.

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