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 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:Store product at 4°C if to be used immediately within two weeks.
For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20°C.
Aliquots are stable at -20°C for 12 months after receipt.
Dilute required amount only prior to immediate use.
Further dilutions can be made in assay buffer.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Note: Applications are based on unconjugated antibody.
仕様
Size：50ug
Host：rabbit
Source Antibody：rat
Grade：Affinity Purified
Purity：Purified by Protein A affinity chromatography.
Form：Supplied as a liquid in PBS, pH 7.4, 0.05% sodium azide, 40% glycerol. Labeled with Biotin. Molar ratio: 1 anitbody: 4 Biotin.
Specificity：Recognizes rat IRS-1 at 160-185kD. Species crossreactivity: Human, mouse and rat.
Isotype：IgG
Calc Applications Abbrev：IP WB
Calc Crossreactivity：Hu Mo Rt
Immunogen：Carboxy terminal 14 amino acid peptide ([C]YASINFQKQPEDRQ) of rat liver IRS-1.
