Detection of Human Phospho-IRS1 (Y612) by Western Blot. Western blot shows lysates of MCF‑7 human breast cancer cell line untreated (-) or treated (+) with 1 µg/mL Insulin for 5 minutes and 300 U/mL CIP for 1 hour. PVDF membrane was probed with 0.2 µg/mL of Mouse Anti-Human Phospho-IRS1 (Y612) Monoclonal Antibody (Catalog # MAB7314) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-IRS1 (Y612) at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Human IRS1 (insulin receptor substrate 1) is a 160-180 kDa substrate that transmits signals from the insulin and IGF-I receptor to downstream signaling modulators. The 1242 amino acid (aa) human IRS1 contains a PH domain (aa 12-115), a PTB domain (aa 160-263) and ten PEST sequences (aa 340-1225). IRS1 may be proteolytically cleaved at Arg656-Val657, generating a 90 kDa and 79 kDa fragment. Upon insulin/IGF-I receptor activation, IRS1 is tyrosine phosphorylated, allowing its association with PI-3 kinase and GRB2. Phosphorylation on Y612 and Y632 is important for IRS1 to fully activate PI3K and translocate GLUT4 in respose to insulin. The sequence surrounding pTyr612 is identical between human and mouse IRS1.
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