Anti-AMPK beta 1 antibody [Y367] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMPK beta 1 antibody [Y367] - BSA and Azide free (AB239804)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder carcinoma tissue sections labeling AMPK beta 1 with purified ab32112 at 1 : 1000 dilution (0.85 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMPK beta 1 antibody [Y367] - BSA and Azide free (AB239804)

Unpurified ab32112 at a 1 : 100 dilution staining AMPK beta 1 in human lung carcinoma, using Immunohistochemistry, Paraffin Embedded Tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-AMPK beta 1 antibody [Y367] - BSA and Azide free (AB239804)

Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling AMPK beta 1 with purified ab32112 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei counterstained with DAPI (blue).

Secondary Only Control : PBS was used instead of the primary antibody as the negative control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AMPK beta 1 antibody [Y367] - BSA and Azide free (AB239804)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling AMPK beta 1 with purified ab32112 at 1 : 1000 dilution (0.85 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-AMPK beta 1 antibody [Y367] - BSA and Azide free (AB239804)

Overlay histogram showing HeLa cells stained with unpurified ab32112 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32112, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-AMPK beta 1 antibody [Y367] - BSA and Azide free (AB239804)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling AMPK beta 1 with purified ab32112 at 1/800 dilution (1 ug/ml) (red). Cells were fixed with 80% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - 0.1% Tween-20. Unlabeled control - Rabbit monoclonal IgG (Black).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

• IP

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Immunoprecipitation - Anti-AMPK beta 1 antibody [Y367] - BSA and Azide free (AB239804)

ab32112 (purified) at 1 : 40 dilution (2ug) immunoprecipitating AMPK beta 1 in NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates.

Lane 1 (input) : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates 10ug

Lane 2 (+) : ab32112 & NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab32112 in NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.

Blocking and diluting buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32112).

All lanes:

Immunoprecipitation - Anti-AMPK beta 1 antibody [Y367] (<a href='/en-us/products/primary-antibodies/ampk-beta-1-antibody-y367-ab32112'>ab32112</a>)

Predicted band size: 30 kDa

Observed band size: 38 kDa

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