Biotin Hamster Anti-Mouse CD119

This SKU will be discontinuing Apr 2024. Suggested alternate SKU is [558771] or for additional support, contact your local applications specialist. Contact Us

BD Pharmingen™ Biotin Hamster Anti-Mouse CD119

Name: Biotin Hamster Anti-Mouse CD119
Brand: BD Pharmingen™
SKU: 550482

Clone 2E2

(RUO)

Expression of cell surface IFN-γRα by BALB/c splenic lymphocytes. RBC-lysed BALB/c spleen cells were preincubated (~15 minutes, 4°C) with purified 2.4G2 antibody [rat anti-mouse CD16 (FcγIII)/CD32 (FcγII); Cat. No. 553142; 1 µg antibody/10e6 cells]. The cells were stained (30 minutes, 4°C) with biotinylated 2E2 antibody (1 µg mAb/10e6 cells; Cat. No. 550482) followed by R-PE-conjugated streptavidin (Cat. No. 554061; 0.015 µg PE-SA/10e6 cells). After washing, the cells were analyzed with a FACScan™ Flow Cytometer. The immunofluorescent staining patterns for cells stained with either biotinylated 2E2 antibody (filled histogram) or Streptavidin-PE (background staining; empty histogram) are shown. The histograms were generated from reanalyzed flow cytometric data files that were gated for events with the light-scattering characteristics of lymphocytes.

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Product Details

Brand: BD Pharmingen™

Alternative Name: Ifngr1; IFN-γ Receptor α; IFN-gamma R alpha; Ifngr; INF-g Receptor; Cd119

Reactivity: Mouse (QC Testing)

Isotype: Armenian Hamster IgG1, κ

Immunogen: Purified preparation of soluble recombinant mouse IFN-γRα chain protein

Application: Flow cytometry (Routinely Tested)

Concentration: 0.5 mg/ml

RRID: AB_393698

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

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

Recommended Assay Procedures

Recommended Assay Procedure:

Immunofluorescent Staining and Flow Cytometric Analysis: The biotinylated form of 2E2 (Cat. No. 550482) can be used for the immunofluorescent staining (≤ 1 µg antibody/10e6 cells) and flow cytometric analysis of normal mouse cells or cell lines to measure their expressed levels of IFN-γRα. It is recommended that the biotin format of this antibody be used in conjunction with streptavidin-phycoerythrin (PE) (Cat No. 554061) in a two-layer staining procedure to amplify immunofluorescent signals. (see figure). An appropriate purified immunoglobulin isotype control is clone A19-3 (Cat. No. 553970).

Note: 2E2 is a nonblocking antibody that can be used for the unobstructed immunofluorescent staining and flow cytometric analysis of cells in systems where the ligand (i.e., IFN-γ) for IFN-γ receptors is present.

Immunoprecipitation: The 2E2 antibody has been reported to be useful for the immunoprecipitation of IFN-γRα chains from lysates of cloned mouse T cells. Please note that this application is not routinely tested at BD Biosciences Pharmingen.

Product Notices

Since applications vary, each investigator should titrate the reagent to obtain optimal results.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.

Companion Products

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO

Size 0.5 mg Cat No. 553142

Biotin Hamster IgG1 κ Isotype Control RUO

Size 0.25 mg Cat No. 553970

PE Streptavidin RUO

Size 0.5 mg Cat No. 554061

550482 Rev. 1

Antibody Details

2E2

The 2E2 antibody recognizes the extracellular region of the 90 kDa alpha chain subunit of the mouse interferon-γ receptor (IFN-γRα; aka, CD119). The functionally active-form of the mouse IFN-γ receptor consists of two (or more) subunits, with IFN-γRα responsible for IFN-γ binding and both the IFN-γRα and IFN-γRβ chains required for the transduction of biologic responses. IFN-γRα is expressed by a variety of cell lines and normal mouse cells (except mature erythrocytes) including T cells, B cells, NK cells, monocytes, neutrophils, fibroblasts, epithelial and endothelial cells. The 2E2 antibody is a non-neutralizing antibody; it does not block the binding of IFN-γ to its receptor. The immunogen used to generate this hybridoma was a purified preparation of soluble recombinant mouse IFN-γRα chain protein.

This antibody is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.

550482 Rev. 1

Format Details

Biotin

Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.

Format: Biotin

550482 Rev.1

Citations & References

Development References (2)

Bach EA, Szabo SJ, Dighe AS, et al. Ligand-induced autoregulation of IFN-gamma receptor beta chain expression in T helper cell subsets.. Science. 1995; 270(5239):1215-8. (Clone-specific: Immunoprecipitation). View Reference
Zola H. Detection of cytokine receptors by flow cytometry. In: Coligan JE, Kruisbeek AM, Margulies DH, Shevach EM, Strober W, ed. Current Protocols in Immunology. New York: Green Publishing Associates and Wiley-Interscience; 1995:6.21.1-6.21.18.

550482 Rev. 1

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.