Anti-Phospho-IRS1 (Ser307) Polyclonal Antibody is a Rabbit antibody targeting Phospho-IRS1 (Ser307). Anti-Phospho-IRS1 (Ser307) Polyclonal Antibody can be used in IHC-P,IHC-Fr,IF,FCM.

IgG

Human (predicted:Mouse,Rat,Dog,Pig,Cow,Horse,Rabbit)

1. Tissue/cell: human kidney carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-phospho-IRS-1 (Ser307) Polyclonal Antibody, Unconjugated (TMAB-01438) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Blank control (blue line): Hela (fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature).
Primary Antibody (green line): Rabbit Anti-phospho-IRS1 (Ser307) antibody (TMAB-01438)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1 μg/test.
3. Blank control: HepG2. Primary Antibody (green line): Rabbit Anti-phospho-IRS1 (Ser307) antibody (TMAB-01438)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: 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-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.

IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg /test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human IRS1 around the phosphorylation site of Ser307

Human

Tumor Associated,Insulin / Insulin-like,Cytoplasmic,Metabolism,Obesity,Heart disease,Diabetes,Cancer,Diabetes associated,Signal transduction