IRS1 (phospho-S307) antibody

• Catalog Number: orb345424
• Category: Antibodies
• Description: IRS1 (phospho-S307) antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IHC, WB
• Reactivity: Human
• Isotype: IgG
• Immunogen: This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal region near amino acids 290-320 of human IRS1 protein.
• Concentration: 1.0 mg/mL
• Dilution range: ELISA: 1:2,000 - 1:10,000, IHC: User Optimized, WB: 1:250 - 1:1,500
• Form/Appearance: Liquid (sterile filtered)
• Purity: 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 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.
• Conjugation: Unconjugated
• UniProt ID: P35568
• NCBI: NP_005535.1
• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: rabbit anti-IRS1 pS307 antibody, IRS-1, IRS 1, HIR
• Note: For research use only
• 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 130 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.
• Expiration Date: 12 months from date of receipt.

Western blot using Biorbyt'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 (p/n orb348669). Lane 2 shows staining of 293 cell lysate prepared from cells serum-starved for 18 h followed by treatment with 5 µg/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 µg 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 1:10000 dilution of IRDye™800 conjugated Gt-a-Rabbit IgG [H&L] MX 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).