Antibody Type: Rabbit Polyclonal

Application: WB,IHC-P,IHC-Fr,IF,FCM

Reactivity: Human,Mouse (predicted:Rat,Dog,Cow,Horse,Sheep)

IgG

Human,Mouse (predicted:Rat,Dog,Cow,Horse,Sheep)

1. Paraformaldehyde-fixed, paraffin embedded (Human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (phospho-AMPK alpha 1 (Ser356)) Polyclonal Antibody, Unconjugated (TMAB-01386) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
2. Paraformaldehyde-fixed, paraffin embedded (Mouse liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (phospho-AMPK alpha 1 (Ser356)) Polyclonal Antibody, Unconjugated (TMAB-01386) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
3. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (phospho-AMPK alpha 1 (Ser356)) Polyclonal Antibody, Unconjugated (TMAB-01386) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. Sample:
Hela (Human) CellLysate at 30 μg
MCF-7 (Human) CellLysate at 30 μg
Primary: Anti-phospho-AMPK alpha 1 (Ser356) (TMAB-01386) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 63 kDa
Observed band size: 63 kDa
5. Sample:
NIH/3T3 (Mouse) Cell Lysate at 30 μg
Primary: Anti-phospho-AMPK alpha 1 (Ser356) (TMAB-01386) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 63 kDa
Observed band size: 63 kDa
6. Blank control (black line): HepG2. Primary Antibody (green line): Rabbit Anti-phospho-AMPK alpha 1 (Ser356) antibody (TMAB-01386)
Dilution: 1 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
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-20°C, 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.

WB,IHC-P,IHC-Fr,IF,FCM

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1ug/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human AMPK alpha 1 around the phosphorylation site of Ser356

Human

5562

Integration of energy metabolism,Response to hypoxia,Metabolism,Integration of energy,Fatty acids,Hypoxia,Fatty acid oxidation,Cancer,Neurodegenerative disease,Amyloid,Other Kinases