Technical Data
IRS-1 (Insulin Receptor Substrate 1) (Biotin)
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.

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 anti-IRS-1 (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.

Included Positive Control:
3T3/A31 cell lysate. Load 20ug of reduced lysate per lane for minigels.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for at least 6 months at -20C. 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.
PabIgGAffinity Purified
50ug4C (-20C Glycerol)Blue IceRatRabbit
As reported
Carboxy terminal 14 amino acid peptide ([C]YASINFQKQPEDRQ) of rat liver IRS-1.
Purified by Protein A affinity chromatography.
Supplied as a liquid in PBS, pH 7.4, 0.05% sodium azide, 40% glycerol. Labeled with Biotin. Molar ratio: 1 anitbody: 4 Biotin.
Recognizes rat IRS-1 at 160-185kD. Species crossreactivity: Human, mouse and rat.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
Lamphere, L. and Lienhard, G.E., Endocrinology 131: 2196, 1992. IRS-1 Review: Myers, M.G., et. al., TIBS 19: 289, 1994.