Phospho-AMPK alpha 2(Ser345) Antibody (YA225)

Phospho-AMPK alpha 2(Ser345) Antibody (YA225) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Phospho-AMPK alpha 2(Ser345).

PRKAA2 is the catalytic subunit of AMP-activated protein kinase (AMPK), a central energy sensor kinase that regulates cellular metabolism. Under ATP depletion, AMPK activates catabolic pathways while inhibiting anabolic processes like lipid/protein synthesis and cell proliferation via direct phosphorylation of metabolic enzymes (e.g., ACACA, ACACB, LIPE) or transcriptional regulators (e.g., CRTC2, FOXO3, p53). It promotes lipolysis by phosphorylating CHKalpha2 and enhances glucose uptake via SLC2A4/GLUT4 translocation, potentially through TBC1D4/AS160 phosphorylation (by similar). AMPK modulates insulin signaling by targeting IRS1, PFKFB2/3 (by similar) and regulates chromatin remodeling via H2B phosphorylation (H2BS36ph) under stress (by similar). As a growth controller, it inhibits mTORC1 by phosphorylating RPTOR/TSC2/WDR24 during nutrient scarcity and initiates autophagy through ATG1/ULK1 activation. It also influences circadian rhythms (by similar), Wnt signaling (by similar), and cell polarity. Additional substrates include CASP6 (apoptosis suppression), ACSS2 (nuclear translocation), and MTFR1L (mitochondrial fission).

Free

P54646

Predicted band size: 62 kDa; Observed band size: 62 kDa

Protein A affinity purified.

Nucleus, Cytoplasm.

Non-conjugated

Phosphorylated

AB_3102528

Neuroscience

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse

WB: 1:500-1:2,000 ;ICC/IF: 1:50-1:200 ;IHC-P: 1:50-1:100

• 
  Western blot analysis of extracts from HEK293 (lane 2(20μg) , HEK293 (lane 3(40μg),using Phospho-AMPK alpha 2 Antibody (HY-P80452). Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of C2C12 cells labeling Phospho-AMPK alpha 2(Ser345) with Phospho-AMPK alpha 2(Ser345) Antibody (HY-P80452) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-AMPK alpha 2(Ser345) (HY-P80452) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of C2C12 cells labeling Phospho-AMPK alpha 2(Ser345) with Phospho-AMPK alpha 2(Ser345) Antibody (HY-P80452) at 1/100dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Phospho-AMPK alpha 2(Ser345)(HY-P80452) at 1/100 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded Rat heart tissue using Phospho-AMPK alpha 2(Ser345) Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80452, 1/100) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded Rat heart tissue using Phospho-AMPK alpha 2(Ser345) Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80452, 1/100) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, ICC/IF, IHC-P

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic phosphopeptide corresponding to residues surrounding Ser345 of Human AMPK alpha 2.AA range:338-352.