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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 CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
The MIH43 monoclonal antibody specifically binds to human B7-H4 (B7 family member, H4) that is encoded by the VTCN1 (V-set domain containing T cell activation inhibitor 1) gene. B7-H4 is a type I membrane glycoprotein with a calculated molecular weight of 30.89 kDa. It is also known as VCTN1, B7h.5, B7S1 (B7 superfamily member 1) or B7X. B7-H4 is a newly discovered member of the B7 family of costimulatory proteins. B7-H4 is not constitutively expressed on peripheral tissues. Its expression can be induced on macrophages, dendritic cells, B cells and T cells. By binding to its putative receptor, the function of B7-H4 was initially reported to be a negative regulator of T cell activation, proliferation and differentiation related to cytokine production and cytotoxic effector cell functions. B7-H4 is reportedly overexpressed in a variety tumors. B7-H4 expression by tumor macrophages appears to play a role in suppression antigen-specific T cell mediated immunity. Recent studies indicate that B7-H4 can also be a positive regulator of T cell responses. B7-H4 can be involved innate immunity as well, eg, by inhibiting neutrophil expansion.
The antibody was conjugated to BD Horizon BUV615 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 615 nm. BD Horizon Brilliant BUV615 can be excited by the ultraviolet laser (355 nm) and detected with a 610/20 filter and a 595 nm LP. Due to the excitation of the acceptor dye by the blue/yellow-green laser line, there may be significant spillover into channels detecting PE-CF594 like emissions (eg, 610/20-nm filter).
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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.
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