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 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 CompBead to ensure that BD Comp beads 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).
Product Notices
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- CF™ is a trademark of Biotium, Inc.
- Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
- BD Horizon Brilliant Violet 605 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
Companion Products
The L203.rMab is a recombinant monoclonal antibody that was derived from L203 hybridoma cells. The L203.rMab specifically recognizes a monomorphic epitope on the extracellular region of human HLA-DR antigens which are human Major Histocompatibility Complex (MHC) Class II antigens. HLA-DR antigens are heterodimers comprised of two different type I transmembrane glycoproteins that are noncovalently-associated. The ~34 kDa HLA-DR alpha (HLA-DRα) chain is encoded by HLA-DRA whereas the ~28 kDa HLA-DR beta (HLA-DRβ) chains are encoded by one of the 4 different HLA-DRB loci (HLA-DRB1,3,4,5) that are located within the Human Leukocyte Antigen (HLA) Complex of chromosome 6. Each chain is comprised of an extracellular region with an IgSF domain, followed by a transmembrane sequence and a short cytoplasmic tail. The L203.rMab antibody recognizes a common determinant that is dependent on the association of HLA-DR alpha and beta chains. HLA-DR is variably expressed on B cells, activated T cells and NK cells, monocytes, macrophages, dendritic cells (DC), Langerhans cells, thymic epithelial cells, and tumor cell lines including B cell lines, myelomas, and some myeloid leukemias. HLA-DR functions in the presentation of peptide antigens to CD4+ T lymphocytes in the generation and regulation of adaptive immune responses. HLA-DR expressed on thymic stromal cells plays a key role in the positive and negative selection of CD4+ T cells during thymopoiesis. Certain HLA-DR alleles, polymorphisms or aberrant expression patterns are associated with susceptibility to diseases including autoimmunity and cancer. L203 antibody binding is reportedly blocked by the L243 monoclonal antibody suggesting that the two antibodies recognize crossreactive or spatially related determinants on HLA-DR.
This antibody is conjugated to BD Horizon™ BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max of 602-nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20-nm filter set. BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605-nm. Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser.
Development References (4)
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Beck B, Dorfel D, Lichtenegger FS, et al. Effects of TLR agonists on maturation and function of 3-day dendritic cells from AML patients in complete remission. J Transl Med. 2011; 9(151):1-14. (Clone-specific: Flow cytometry). View Reference
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Finn OJ, Levy R. Multiple HLA-DR antigens: detection with monoclonal antibodies and translation in vitro.. Proc Natl Acad Sci USA. 1982; 79(8):2658-62. (Clone-specific: Immunoprecipitation). View Reference
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Lampson LA, Levy R. Two populations of Ia-like molecules on a human B cell line.. J Immunol. 1980; 125(1):293-9. (Immunogen: Blocking, Immunoprecipitation, Radioimmunoassay). View Reference
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Suni MA, Picker LJ, Maino VC. Detection of antigen-specific T cell cytokine expression in whole blood by flow cytometry.. J Immunol Methods. 1998; 212(1):89-98. (Clone-specific: Blocking, Functional assay, Inhibition). View Reference
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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.
