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Immunofluorescence analysis of AMPK alpha 1 + 2 [pT172] 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 AMPK alpha 1 + 2 [pT172] Rabbit Polyclonal Antibody (441150G) at 1ug/mL 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 (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 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.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human AMPK that contains threonine 172. The sequence is conserved in human, mouse and rat.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||1 µg/ml|
|Immunofluorescence (IF)||1 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:50|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
The protein encoded by this gene is a catalytic subunit of the AMP-activated protein kinase (AMPK). AMPK is a heterotrimer consisting of an alpha catalytic subunit, and non-catalytic beta and gamma subunits. AMPK is an important energy-sensing enzyme that monitors cellular energy status. In response to cellular metabolic stresses, AMPK is activated, and thus phosphorylates and inactivates acetyl-CoA carboxylase (ACC) and beta-hydroxy beta-methylglutaryl-CoA reductase (HMGCR), key enzymes involved in regulating de novo biosynthesis of fatty acid and cholesterol. Studies of the mouse counterpart suggest that this catalytic subunit may control whole-body insulin sensitivity and is necessary for maintaining myocardial energy homeostasis during ischemia.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
5'-AMP-activated protein kinase; 5'-AMP-activated protein kinase catalytic subunit alpha-1; 5'-AMP-activated protein kinase catalytic subunit alpha-2; 5'-AMP-activated protein kinase, catalytic alpha-1 chain; 5'-AMP-activated protein kinase, catalytic alpha-2 chain; ACACA kinase; acetyl-CoA carboxylase kinase; alpha 2 catalytic subunit; alpha-1; AMP -activate kinase alpha 1 subunit; AMP-activated; AMP-activated protein kinase; AMP-activated protein kinase alpha 2 catalytic subunit; AMP-activated protein kinase alpha-2 subunit variant 2; AMP-activated protein kinase alpha-2 subunit variant 3; AMP-activated protein kinase alpha-2 variant B; AMP-activated protein kinase, alpha 1 catalytic subunit; AMP-activated protein kinase, catalytic, alpha-1; AMPK; AMPK alpha 1; AMPK alpha 2; AMPK subunit alpha-1; AMPK subunit alpha-2; AMPK-alpha-2 chain; AMPK2; AMPKa1; catalytic; catalytic alpha-1 chain; catalytic alpha-2 chain; HMGCR kinase; hydroxymethylglutaryl-CoA reductase kinase; pAMPK; PRKAA; PRKAA2; protein kinase; protein kinase, AMP-activated, alpha 1 catalytic subunit; protein kinase, AMP-activated, alpha 2 catalytic subunit; tau-protein kinase PRKAA1
2310008I11Rik; A830082D05; AI194361; AI450832; AL024255; AMPK; AMPK1; AMPK2; AMPKa1; AMPKa2; AMPKalpha1; AMPKalpha2; C130083N04Rik; PRKAA; PRKAA1; PRKAA2