Phospho-AMPK alpha-1 (Ser496+Ser502) Rabbit Polyclonal Antibody

• Catalog Number: orb5696
• Category: Antibodies
• Description: Phospho-AMPK alpha-1 (Ser496+Ser502) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P
• Predicted Reactivity: Rat
• Reactivity: Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from mouse AMPK alpha-1 around the phosphorylation site of Ser496+Ser502 SG(p-S)IS
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=2ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 64 kDa
• Target: PRKAA1
• UniProt ID: Q5EG47
• RRID: AB_10935854
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: AMPK alpha 2(S496+S502); AMPK alpha 1(phospho S496
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (Black line): Mouse spleen (Black). Primary Antibody (green line): Rabbit Anti-phospho-AMPK alpha-1 (Ser496+Ser502) antibody (orb5696), dilution: 3 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647, dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 10000 events was performed.

Blank control: Mouse spleen. Primary Antibody (green line): Rabbit Anti-AMPK antibody (orb5696), dilution: 2 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, dilution: 1 µg/Test. Protocol, The cells were fixed with 70% ethanol (10 min at room temperature) and then were incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (Mouse brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-AMPK alpha-1 (Ser496+Ser502)) Polyclonal Antibody, Unconjugated (orb5696) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-AMPK alpha-1 (Ser496+Ser502)) Polyclonal Antibody, Unconjugated (orb5696) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.