This affinity-purified antibody is directed against the phosphorylated form of human IRS1 protein at the pS307 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity occurs against human IRS1 pS307 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human IRS1 is minimal by ELISA. A BLAST analysis was used to suggest cross reactivity with IRS1 from human, mouse, rat, dog and vervet monkey based on 100 % homology with the immunizing sequence. Partial reactivity is also expected against IRS1 from pig (94 %), bovine (94 %) and chicken (88 %) sources. Reactivity of this antibody with IRS1 from other species is unknown.
Purification
Immunoaffinity chromatography.
Immunogen
This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids 298-316 of human IRS1 protein.
IRS1
Reactivity: Human, Mouse, Rat
WB, IHC, ELISA, IP, DB
Host: Rabbit
Polyclonal
unconjugated
Application Notes
This affinity purified antibody has been tested for use in ELISA (1: 2,000-1: 10,000) andwestern blot (1: 250-1: 1,500). Specific conditions for reactivity should be optimized by theend user. Expect a band approximately 180 kDa in size corresponding to phosphorylatedIRS1 protein by western blotting in the appropriate cell lysate or extract. Minimal reactivityis observed against the non-phosphorylated form of the immunizing peptide. This antibodyis phospho specific for pS307 of IRS1 protein. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Restrictions
For Research Use only
Concentration
0.5 mg/mL (by UV absorbance at 280 nm)
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 with 0.01 % (w/v) Sodium Azide as preservative.
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Avoid repeated freezing and thawing.
Storage
4 °C
Storage Comment
Store undiluted at 2-8 °Cfor one month or (in aliquots) at -20 °C for longer.
Insulin Receptor Substrate 1 (IRS1) acts as a signaling molecule for IL-4, insulin and insulin-like growth factor-I (IGF-I) receptors. When phosphorylated by the insulin receptor, IRS1 binds specifically to various cellular proteins containing SH2 domains such as phosphatidylinositol 3-kinase p85 subunit or GRB2. When bound, IRS1 typically activates phosphatidylinositol 3-kinase p85 subunit. IRS1 interacts with both the NPXY motif of tyrosine-phosphorylated IGF1R and the INSR through the PTB domain. Serine phosphorylation of IRS1 is a mechanism for insulin resistance. Ser-312 phosphorylation inhibits insulin action through disruption of IRS1 interaction with the insulin receptor. Ser-307 phosphorylation is involved in the mechanism of insulin resistance in type 2 diabetes. Polymorphisms in IRS1 may be involved in the etiology of non-insulin-dependent diabetes mellitus (NIDDM).Synonyms: IRS-1, Insulin receptor substrate 1