Detection of Human Phospho-IRS1 (S616) 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 1 µg/mL of Mouse Anti-Human Phospho-IRS1 (S616) Monoclonal Antibody (Catalog # MAB39783) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-IRS1 (S616) 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 150-180 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. When PI-3 kinase and ERK1/2 are activated, insulin or angiotensin II can also stimulate phosphorylation on Ser616. The aa sequence surrounding pSer616 is identical between human, mouse and rat IRS-1.
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