Rabbit anti-Human IRS1 Polyclonal Antibody | anti-IRS1 antibody

Anti-Human IRS1 Phospho-Ser307 Antibody

Cat. Num. AAA4157016

• Reactivity: Human
• Applications: Western Blot, Immunohistochemistry, ELISA
• Purity: Affinity Purified
• Synonyms: IRS1; Polyclonal Antibody; Anti-Human IRS1 Phospho-Ser307 Antibody; Human IRS1; anti-IRS1 antibody

• Host: Rabbit
• Reactivity: Human
• Clonality: Polyclonal
• Isotype: IgG
• Specificity: This affinity-purified antibody is directed against the phosphorylated form of human IRS1 protein at the pS307 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross-adsorbed ELISA 1:2,000-1:10,000 WESTERN BLOT 1:250-1:1,500 IMMUNOHISTOCHEMISTRY User Optimized OTHER APPLICATIONS User Optimized Figure 1. Western blot using Rockland's Affinity Purified anti-IRS1 pS307 antibody shows detection of a band at ~180 kDa believed to represent phosphorylated IRS1 (arrowhead). Lane 1 shows staining of human 293 cell lysate. Lane 2 shows staining of 293 cell lysate prepared from cells serum-starved for 18 h followed by treatment with 5 靏/ml of anisomysin for 30 min. The pronounced staining of the band at 180 kDa is not seen when the antibody was pre-incubated with immunizing peptide prior to reaction (data not shown). The identity of the intensely reactive bands at ~70 kD in both lane 1 and 2 is unknown, although these bands were also competed out by pre-incubation with the immunizing peptide. Approximately 25 靏 of each lysate was separated on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred onto nitrocellulose. After blocking with 5% goat serum, 0.5% BLOTTO in PBS, the membrane was probed with the primary antibody diluted to 1:250. Reaction occurred overnight at 4� C followed by washes and reaction with a 1:10,000 dilution of IRDye� conjugated Gt-a-Rabbit IgG [H&L] MX (611-132-122) for 45 min at room temperature (800 nm channel, green). Molecular weight estimation was made by comparison to prestained MW markers in lane M (700 nm channel, red). IRDye� fluorescence image was captured using the Odyssey� Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results. M 1 2 170 130 100 55 40 33 Page 2 of 2 e-mail: [email protected] Visit our website at: www.rockland-inc.com against the non-phosphorylated form of the immunizing peptide. Reactivity occurs against human IRS1 pS307 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human IRS1 is minimal by ELISA. A BLAST analysis was used to suggest cross reactivity with IRS1 from human, mouse, rat, dog and vervet monkey based on 100% homology with the immunizing sequence. Partial reactivity is also expected against IRS1 from pig (94%), bovine (94%) and chicken (88%) sources. Reactivity of this antibody with IRS1 from other species is unknown.
• Purity/Purification: Affinity Purified
• Form/Format: Liquid; PBS, pH 7.4 with 0.05% sodium azide.
• Concentration: 1ug/ul (varies by lot)
• Sequence Length: 518

• Applicable Applications for anti-IRS1 antibody: Western Blot (WB), Immunohistochemistry (IHC), ELISA (EIA)
• Application Notes: This affinity purified antibody has been tested for use in ELISA and western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 180 kDa in size corresponding to phosphorylated IRS1 protein by western blotting in the appropriate cell lysate or extract. Minimal reactivity is observed against the non-phosphorylated form of the immunizing peptide. This antibody is phospho specific for pS307 of IRS1 protein.
• Immunogen: This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids 298-316 of human IRS1 protein.
• Preparation and Storage: This product is stable for several weeks at 4 degree C as an undiluted liquid. Dilute only prior to immediate use.; For extended storage, aliquot contents and freeze at-20 degree C or below. Avoid cycles of freezing and thawing. Expiration date is one (1) year from date of receipt.

Testing Data

Related Product Information for anti-IRS1 antibody

Insulin Receptor Substrate 1 (IRS1) acts as a signaling molecule for IL-4, insulin and insulin-like growth factor-I (IGF-I) receptors. When phosphorylated by the insulin receptor, IRS1 binds specifically to various cellular proteins containing SH2 domains such as phosphatidylinositol 3-kinase p85 subunit or GRB2. When bound, IRS1 typically activates phosphatidylinositol 3-kinase p85 subunit. IRS1 interacts with both the NPXY motif of tyrosine-phosphorylated IGF1R and the INSR through the PTB domain. Serine phosphorylation of IRS1 is a mechanism for insulin resistance. Ser-312 phosphorylation inhibits insulin action through disruption of IRS1 interaction with the insulin receptor. Ser-307 phosphorylation is involved in the mechanism of insulin resistance in type 2 diabetes. Polymorphisms in IRS1 may be involved in the etiology of non-insulin-dependent diabetes mellitus (NIDDM).

NCBI and Uniprot Product Information

• NCBI GI #: 958211643
• NCBI Official Full Name: IRS1, partial
• Protein Family: Isoleucine--tRNA ligase