Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
Companion Products
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.
Development References (6)
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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
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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
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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.
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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
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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
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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
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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.