Detection of Human Phospho-IRS1 (Y1179) 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 (Y1179) Monoclonal Antibody (Catalog # MAB7455) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-IRS1 (Y1179) at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Phospho-IRS1 (Y1179) in MCF‑7 Human Cell Line.
IRS1 phosphorylated at Y1179 was detected in immersion fixed MCF‑7 human breast cancer cell line using Mouse Anti-Human Phospho-IRS1 (Y1179) Monoclonal Antibody (Catalog # MAB7455) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
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 intermediate between the insulin and IGF-I receptor,
and downstream signaling modulators. Upon insulin/IGF-I receptor
activation, IRS1 is tyrosine phosphorylated, allowing its association with PI-3
kinase and GRB2. Human IRS1 is 1242 amino acids (aa) in length. It
contains a PH (pleckstrin homology) domain (aa 12-115), followed by a PTB
(phosphotyrosine-binding) domain (aa 160-263) and ten PEST (Pro/Glu/Ser/Thr) regions
(aa 340-1225). IRS1 may be proteolytically cleaved at Arg656-Val657,
generating a 90 kDa and 79 kDa fragment. Insulin receptor-mediated
phosphorylation of human IRS1 on Y1179, or its murine equivalent, Y1172,
mediates IRS-1 interaction with SH2 domains such as that of the Src family
kinase, Fyn, or the tyrosine phosphatase, SHP-2.
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