Technical Data
I8700
IRS-1 (Insulin Receptor Substrate 1)
Description:
IRS-1 is one of the major substrates of insulin receptor kinase. IRS-1 protein contains a conserved pleckstrin homology (PH) domain located at its amino terminus, adjacent to a phospho-tyrosine binding (PTB) domain. IRS-1 contains multiple tyrosine phosphorylation motifs that serve as docking sites for SH2 domain–containing proteins (Grb2, p85, SHP-2, Fyn and others) and which mediate the metabolic and growthpromoting functions of insulin. IRS-1 also contains over 30 potential Ser/Thr phosphorylation sites for kinases, such as PKA, PKC and MAPK. Once phosphorylated by insulin receptor, IRS-1 recruits the downstream components that mediate the insulin signaling.

Applications:
Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested.

Recommended Dilution:
Western Blot: 0.5-2ug/ml detects IRS-1 in RIPA lysates from 3T3/A31 mouse fibroblasts. 3T3/A31 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with I8700 (0.5ug/ml). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system. Previous lots detected IRS-1 in RIPA lysates of rat L6 myoblasts. In cells that contain high levels of IRS-1, this lot may work optimally at 0.1-0.5ug/ml.
Immunoprecipitation: 4ug immunoprecipitates IRS-1 from 500ug of 3T3/A31 RIPA lysate
Optimal dilutions to be determined by the researcher.

Positive Control:
Fibroblast Cell, 3T3/A31, L6 myoblasts.
Recommended Secondary Antibodies
I1904-39: IgG, X-Adsorbed (HRP) Pab Gt xRb
I1904-40A: IgG, H&L, X-Adsorbed (HRP) Pab Gt xRb
I1904-46Q: IgG, H&L (HRP) Pab Gt xRb
Storage and Stability:
May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
100ug4°C (-20°C Glycerol)Blue IceRatRabbit
Concentration:
~0.25mg/ml
Immunogen:
Synthetic peptide corresponding to the Carboxy terminal 14aa, (C)YASINFQKQPEDRQ of rat liver IRS-1.
Purity:
Purified by immunoaffinity chromatography.
Form
Supplied as a liquid in 0.2M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.1mM EDTA, 5mg/ml BSA, 0.05% sodium azide.
Specificity:
Recognizes rat IRS-1 at 160-185kD. Species crossreactivity: human and mouse
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Lamphere, L. and Lienhard, G.E., Endocrinology 131: 2,196, 1992. 2. Takano, A., et al., Mol. Cell. Biol. 21: 5,050, 2001. 3. Lingamanaidu, V., et al., J. Biol. Chem. 276: 3,543, 2001. 4. Myers, M.G., et al., TIBS 19: 289, 1994.