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BUV661 Mouse Anti-Human HLA-DP

BD OptiBuild™ BUV661 Mouse Anti-Human HLA-DP

Clone B7/21 (also known as B7/21.49; B7/21 (IgG1))

(RUO)
Product Details
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BD OptiBuild™
HLADP; HLA-DPαβ; DPA and DPB; DPα and DPβ; DPαβ
Human (Tested in Development)
Mouse BALB/c IgG1, κ
Human NALM6 Leukemia Cells
Flow cytometry (Qualified)
0.2 mg/ml
AB_2875024
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. BD Horizon Brilliant Ultraviolet 661 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.
750943 Rev. 3
Antibody Details
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B7/21

The B7/21 monoclonal antibody specifically recognizes HLA-DP antigens which are human Major Histocompatibility Complex (MHC) Class II antigens. HLA-DP antigens are heterodimers comprised of two different type I transmembrane glycoproteins that are noncovalently-associated. The ~34 kDa HLA-DP alpha (HLA-DPα) and ~26-28 kDa HLA-DP beta (HLA-DPβ) chains are encoded by HLA-DPA1 and HLA-DPB1 genes, respectively, that are located within the Human Leukocyte Antigen (HLA) Complex of chromosome 6. The B7/21 antibody recognizes a monomorphic determinant present on cells expressing HLA-DP1, -DP2, -DP3, -DP4, and -DP5 and does not reportedly bind to HLA-DR or HLA-DQ Class II antigens. HLA-DP is variably expressed on B cells, monocytes, macrophages, dendritic cells (DC), activated T cells, and tumor cell lines including B cell lines, myelomas, and some myeloid leukemias. HLA-DP functions in the presentation of peptides derived from extracellular antigens to CD4+ T cells.

       The original B7/21 hybridoma was derived from hybridization of mouse 5194/SXXOBU1 myeloma cells with spleen cells from BALB/c mice immunized with NALM6 human leukemia cells and secreted IgG3, κ antibody. An immunoglobulin switch-variant B7/21 hybridoma, also known as B7/21.49 or B7/21 (IgG1), was subsequently isolated that secreted lgG1 antibody with the same specificity as the original IgG3 antibody. The purified switch-variant B7/21 IgG1 antibody blocks the staining of HLA-DP-positive cells by fluorescent B7/21 IgG3 antibody.  The B7/21 IgG1 antibody is reportedly not cytotoxic in the presence of complement.

The antibody was conjugated to BD Horizon™ BUV661 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 661-nm. BD Horizon Brilliant BUV661 can be excited by the ultraviolet laser (355 nm) and detected with a 670/25 filter and a 630 nm LP.  Due to cross laser excitation of this dye, there may be significant spillover into channels detecting APC-like emissions (eg, 670/25-nm filter).

    

Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV661 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV661 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone-specific compensation controls when using these reagents.

750943 Rev. 3
Format Details
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BUV661
The BD Horizon Brilliant™ Ultraviolet 661 (BUV661) 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 660-nm. BUV661, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 660-nm (e.g., 670/25 bandpass filter). The acceptor dye can be excited by the Red (628–640-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.
BUV661
Ultraviolet 355 nm
350 nm
660 nm
750943 Rev.3
Citations & References
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Development References (2)

  1. Lennon-Duménil AM, Barbouche MR, Vedrenne J, et al. Uncoordinated HLA-D gene expression in a RFXANK-defective patient with MHC class II deficiency.. J Immunol. 2001; 166(9):5681-7. (Clone-specific). View Reference
  2. Strobl H, Bello-Fernandez C, Riedl E, et al. flt3 ligand in cooperation with transforming growth factor-beta1 potentiates in vitro development of Langerhans-type dendritic cells and allows single-cell dendritic cell cluster formation under serum-free conditions.. Blood. 1997; 90(4):1425-34. (Clone-specific). View Reference
750943 Rev. 3

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.