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PE Mouse Anti-Human CD119
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PE Mouse Anti-Human CD119
Flow cytometric analysis of CD119 expression on human PBMC isolated by Lymphoprep (Nycomed) density centrifugation. Whole blood was stained either with PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; open histogram) or PE Mouse Anti-Human CD119 (Cat. No. 558934/558937/557531; filled histogram) The fluorescence histograms were derived from gated events with the forward and side light-scattering characteristics of viable cells.
Flow cytometric analysis of CD119 expression on human PBMC isolated by Lymphoprep (Nycomed) density centrifugation. Whole blood was stained either with PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; open histogram) or PE Mouse Anti-Human CD119 (Cat. No. 558934/558937/557531; filled histogram) The fluorescence histograms were derived from gated events with the forward and side light-scattering characteristics of viable cells.
Product Details
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BD Pharmingen™
IFN- gamma receptor alpha chain
Human (QC Testing)
Mouse IgG1, κ
Human IFN-γRα
Flow cytometry (Routinely Tested)
0.2 mg/ml
VI C-110
3459
AB_397163
Aqueous buffered solution containing ≤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.

Recommended Assay Procedures

Immunofluorescent staining and flow cytometric analysis: PE Mouse Anti-Human CD119 (Cat. No. 558934) can be used for the immunofluorescent staining (≤ 1 µg antibody/10[6] cells) and flow cytometric analysis of the levels of membrane IFN-γRα expressed by human cell lines or human lymphoid cells. An appropriate R-PE conjugated immunoglobulin isotype control is the PE Mouse IgG1, κ Isotype Control (Cat. No. 554680).

Since GIR-208 is a neutralizing antibody, it competes with IFN-γ for binding to its receptor. Therefore, the use of the GIR-208 antibody for

immunofluorescent staining and flow cytometric analysis in systems where the natural ligand of the receptor is present may give an underestimation of IFN-γRα chain expression. Based on our testing results, the presence of recombinant human IFN-γ protein at levels above 200 ng/ml is sufficient to completely inhibit the binding of the GIR-208 antibody (at 0.06 µg/10[6] cells). In such cases, it is recommended that the investigator uses a non-neutralizing antibody to detect IFN-γRα chain expression by flow cytometry, such as PE Mouse Anti-Human CD119 (Cat. No. 558937).

WB: The GIR-208 antibody has been reported to be useful for Western blotting.  Please note that this application is not routinely tested at BD Biosciences Pharmingen.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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.
558934 Rev. 3
Antibody Details
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GIR-208

The GIR-208 antibody recognizes the extracellular region of CD119 which is also known as the alpha chain subunit (80-95 kDa glycoprotein) of the human interferon-γ receptor (IFN-γRα).  The functionally active-form of the human IFN-γ receptor consists of two (or more) subunits, with IFN-γRα responsible for IFN-γ binding and both the IFN-γRα and β chains required for the transduction of biologic responses.  The IFN-γ receptor α chain (CD119) is expressed on the surface of most human cells (except mature erythrocytes) including monocytes, macrophages, T cells, B cells, NK cells, neutrophils, fibroblasts, epithelial cells, and endothelium.  Binding of 125I-labeled GIR-208 antibody to IFN-γRα+ cells is reported to be specifically inhibited in the presence of excess IFN-γ.  GIR-208 does not cross react with IFN-γ as tested by ELISA. The ability of this antibody to bind to IFN-γ receptors of species other than human has not been determined. The immunogen used to generate this hybridoma was human IFN-γRα purified from human placenta.  The GIR- 208 has been reported to block the binding of 125I-human IFN-γ to IFN-γRα+ cells as well as purified, soluble human IFN-γRα. GIR-208 is a neutralizing antibody that has been shown to neutralize the anti-viral activity of IFN-γ on WISH cells in a dose-dependent fashion.

558934 Rev. 3
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.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
558934 Rev.3
Citations & References
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Development References (6)

  1. Bach EA, Aguet M, Schreiber RD. The IFN gamma receptor: a paradigm for cytokine receptor signaling. Annu Rev Immunol. 1997; 15:563-591. (Biology). View Reference
  2. Gumina RJ, Freire-Moar J, DeYoung L, Webb DR, Devens BH. Transduction of the IFN-gamma signal for HLA-DR expression in the promonocytic line THP-1 involves a late-acting PKC activity. Cell Immunol. 1991; 138(2):265-279. (Clone-specific: Neutralization). View Reference
  3. Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:818-821.
  4. Peyman JA, Hammond GL. Localization of IFN-gamma receptor in first trimester placenta to trophoblasts but lack of stimulation of HLA-DRA, -DRB, or invariant chain mRNA expression by IFN-gamma. J Immunol. 1992; 149(8):2675-2680. (Biology). View Reference
  5. Sheehan KC, Calderon J, Schreiber RD. Generation and characterization of monoclonal antibodies specific for the human IFN-gamma receptor. J Immunol. 1988; 140(12):4231-4237. (Immunogen: Neutralization, Western blot). View Reference
  6. Valente G, Ozmen L, Novelli F. Distribution of interferon-gamma receptor in human tissues. Eur J Immunol. 1992; 22(9):2403-2412. (Biology). View Reference
View All (6) View Less
558934 Rev. 3

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