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

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

IRS1

Insulin receptor substrate 1

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

May mediate the control of various cellular processes by insulin. When phosphorylated by the insulin receptor binds specifically to various cellular proteins containing SH2 domains such as phosphatidylinositol 3-kinase p85 subunit or GRB2. Activates phosphatidylinositol 3-kinase when bound to the regulatory p85 subunit.