Technical Data
IRS-1, phosphorylated (Ser307) (Insulin Receptor Substrate 1) (Glycerol Free)
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 growth-promoting 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. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000-1:2000 detects phospho-IRS-1 (Ser307) in RIPA lysates from serum starved HEK 293 cells treated with 5ug/ml anisomycin for 30 minutes, but not in untreated cells. Optimal dilutions to be determined by the researcher.

Immunoblot Analysis:
HEK 293 cell lysates untreated and anisomycin stimulated were resolved by electrophoresis, transferred to nitrocellulose and probed with antiphospho-IRS-1 (Ser307) (1:1000 dilution) or anti-IRS-1 (1:1000 dilution). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.

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 12 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.
200ul-20CBlue IceRatRabbit
Not determined
Synthetic peptide corresponding to aa301-315 (ESITATpSPASMVGGK) of phospho-IRS-1 (Ser307) (KLH coupled).
Supplied as a liquid in TBS, 0.05% sodium azide. Glycerol free.
Recognizes phospho-IRS-1 (Ser307), Mr 170kD. Species Crossreactivity: Human, mouse and rat. Species sequence homology: mouse, human, rat and monkey:100%; chicken: 12/15.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Aguirre, V., et al., J. Biol. Chem. 275: 9,047-9,054 (2000). 2. Gao, Z., et al., J. Biol. Chem. 277: 48,115-48,121 (2002). 3. Rui, L., et al., J. Clin. Invest. 107: 181-189 (2001).