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PE Mouse anti-IRS-1 (pY896)
PE Mouse anti-IRS-1 (pY896)

Analysis of IRS-1 (pY896) in transformed human epithelial cells.  Serum-starved 293 fetal kidney cells were either stimulated for 2 minutes with 100 nM  IGF-I (Cat. No. 354037, shaded histogram) or unstimulated (open histogram).  The cells were fixed (BD Phosflow™ Fix Buffer I, Cat. No. 557870) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with PE Mouse anti-IRS-1 (pY896, Cat. No. 558440).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Analysis of IRS-1 (pY896) in transformed human epithelial cells.  Serum-starved 293 fetal kidney cells were either stimulated for 2 minutes with 100 nM  IGF-I (Cat. No. 354037, shaded histogram) or unstimulated (open histogram).  The cells were fixed (BD Phosflow™ Fix Buffer I, Cat. No. 557870) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with PE Mouse anti-IRS-1 (pY896, Cat. No. 558440).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Product Details
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BD Phosflow™
Human (QC Testing), Mouse, Rat (Predicted)
Mouse BALB/c IgG2a, κ
Phosphorylated Human IRS-1 Peptide
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
AB_647216
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
558440 Rev. 2
Antibody Details
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K9-211

The IRS (Insulin Receptor Substrate) proteins IRS-1, IRS-2, IRS-3, and IRS-4 are major substrates of the insulin receptor and the insulin-like growth factor-1 (IGF-1) receptor tyrosine kinases.  IRS proteins contain an N-terminal pleckstrin homology (PH) domain, a phosphotyrosine-binding (PTB) domain, and multiple tyrosine phosphorylation sites in the C-terminus.  The IRS-1 protein is widely expressed and, along with IRS-2, mediates somatic growth and carbohydrate metabolic responses to insulin.  Following insulin receptor ligation, IRS-1 binds to the juxtamembrane region of the receptor via the PH and PTB domains and is tyrosine phosphorylated, which facilitates its interaction with SH2 domain-containing signaling proteins.  Specifically, the phosphorylated tyrosine 896 (pY896) of human IRS-1 is a major binding site for the GRB2 (Growth-factor Receptor-Bound protein 2) adaptor protein.  After IRS-1 activation, negative and positive feedback regulates dephosphorylation of its tyrosine sites, which ultimately regulates the magnitude and/or duration of the downstream pleiotropic responses to insulin and IGF-1.

The K9-211 monoclonal antibody recognizes pY896 of human IRS-1.  The orthologous phosphorylation sites of mouse and rat IRS-1 are Y891 and Y895, respectively.

558440 Rev. 2
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
558440 Rev.2
Citations & References
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Development References (5)

  1. Burks DJ, White MF. IRS proteins and β-cell function. Diabetes. 2001; 50(S1):S140-S145. (Biology). View Reference
  2. Gual P, Le Marchand-Brustel Y, Tanti JF. Positive and negative regulation of insulin signaling through IRS-1 phosphorylation. Biochimie. 2005; 87:99-109. (Biology). View Reference
  3. Paz K, Liu YF, Shorer H, et al. Phosphorylation of insulin receptor substrate-1 (IRS-1) by protein kinase B positively regulates IRS-1 function. J Biol Chem. 1999; 274(40):28816-28822. (Biology). View Reference
  4. Ward CW, Gough KH, Rashke M, Wan SS, Tribbick G, Wang J. Systematic mapping of potential binding sites for Shc and Grb2 SH2 domains on insulin receptor substrate-1 and the receptors for insulin, epidermal growth factor, platelet-derived growth factor, and fibroblast growth factor. J Biol Chem. 1996; 271(10):5603-5609. (Biology).
  5. White MF. IRS proteins and the common path to diabetes. Am J Physiol Endocrinol Metab. 2002; 283:E413-E422. (Biology). View Reference
View All (5) View Less
558440 Rev. 2

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.