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BUV563 Rat Anti-Human HLA-B
Product Details
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BD OptiBuild™
HLA-B; HLAB
Human (Tested in Development)
Rat IgG1, κ
Normal Human Lymphocytes
Flow cytometry (Qualified)
0.2 mg/ml
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 to the dye under optimum conditions that minimize unconjugated dye and antibody.

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 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).

Product Notices

  1. 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.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. CF™ is a trademark of Biotium, Inc.
  10. BD Horizon Brilliant Ultraviolet 563 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
752622 Rev. 1
Antibody Details
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YTH 76.3.rMAb

The YTH 76.3rMAb is a recombinant monoclonal antibody that specifically recognizes a monomorphic epitope on the extracellular region of the Human Leukocyte Antigen-B (HLA-B) heavy chain. This major histocompatibility complex (MHC) class I antigen is comprised of a polymorphic HLA-B heavy chain, a ~45 kDa type I transmembrane glycoprotein encoded by HLA-B, that is noncovalently associated with the invariant ~12 kDa Beta-2 (β2)-microglobulin light chain encoded by B2M. Hundreds of HLA-B alleles have been reported. HLA-B is expressed on most nucleated cells and leucocytes including specialized antigen presenting cells such as thymic epithelial cells and dendritic cells. HLA-B plays a major role in the MHC-restricted presentation of small peptides, including those derived from self or foreign antigens, that can be bound by TCR expressed on CD8+ thymocytes or T cells leading to either immune tolerance, the maturation of naïve CD8+ T cells, or the activation and differentiation of CD8+ cytotoxic effector T cells or memory CD8+ T cells. HLA-B also functions as a ligand for regulatory CD8 coreceptor molecules and some CD158 molecules that serve as regulatory MHC class I antigen receptors expressed by CD8+ T cells and NK cells. The YTH 76.3rMAb is derived from the hybridoma clone YTH 76.3, with Rat IgG2a, κ isotype. YTH 76.3rMAb has variable region sequences from the original YTH 76.3 clone appended to Rat IgG1 and kappa constant region sequences.

The antibody was conjugated to BD Horizon™ BUV563 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 which has an Ex Max of 348 nm and an acceptor dye. The tandem has an Em Max at 563 nm. BD Horizon BUV563 can be excited by the 355 nm ultraviolet laser. On instruments with a 561 nm Yellow-Green laser, the recommended bandpass filter is 585/15 nm with a 535 nm long pass to minimize laser light leakage. When BD Horizon BUV563 is used with an instrument that does not have a 561 nm laser, a 560/40 nm filter with a 535 nm long pass may be more optimal. Due to the excitation and emission characteristics of the acceptor dye, there may be spillover into the PE and PE-CF594 detectors. However, the spillover can be corrected through compensation as with any other dye combination.

752622 Rev. 1
Format Details
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BUV563
The BD Horizon Brilliant™ Ultraviolet 563 (BUV563) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 564-nm. BUV563, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 560-nm (e.g., a 560/40 or a 585/15-nm bandpass filter). The acceptor dye can be excited by the Blue (488-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV563
Ultraviolet 355 nm
350 nm
564 nm
752622 Rev.1
Citations & References
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Development References (5)

  1. Burrone OR, Kefford RF, Gilmore D, Milstein C. Stimulation of HLA-A,B,C by IFN-alpha. The derivation of Molt 4 variants and the differential expression of HLA-A,B,C subsets.. EMBO J. 1985; 4(11):2855-60. (Clone-specific: Flow cytometry). View Reference
  2. Hale G, Clark M, Waldmann H. Therapeutic potential of rat monoclonal antibodies: isotype specificity of antibody-dependent cell-mediated cytotoxicity with human lymphocytes.. J Immunol. 1985; 134(5):3056-61. (Immunogen). View Reference
  3. Margulies DH, Natarajan K, Rossjohn J, McCluskey J. Major Histocompatibility Complex and Its Proteins. In: Paul WE. Paul WE, ed. Fundamental Immunology 7th Edition. Philadelphia: Lippincott Williams & Wilkins; 2013:487-523.
  4. Rojas RE, Balaji KN, Subramanian A, Boom WH. Regulation of human CD4(+) alphabeta T-cell-receptor-positive (TCR(+)) and gammadelta TCR(+) T-cell responses to Mycobacterium tuberculosis by interleukin-10 and transforming growth factor beta.. Infect Immun. 1999; 67(12):6461-72. (Clone-specific: Flow cytometry). View Reference
  5. Schmidt H, Gekeler V, Haas H, et al. Differential regulation of HLA class I genes by interferon.. Immunogenetics. 1990; 31(4):245-52. (Biology). View Reference
View All (5) View Less
752622 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.