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.
- 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.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The UCH-B1 (also known as, UCHB1) monoclonal antibody specifically recognizes the heavy chain constant region (Cµ) of human Immunoglobulin M (IgM) that is encoded by IGHM (immunoglobulin heavy constant mu). It does not crossreact with other immunoglobulin heavy or light chain isotypes. An intracytoplasmic form of IgM is expressed by pre-B cells whereas immature and a portion of mature B cells, including naive and memory B cells, express cell surface IgM. These forms of IgM can also be expressed by cells from some leukemias or lymphomas. Cell surface IgM is comprised of two type I transmembrane heavy chain glycoproteins (Igµ heavy chains) that are paired with immunoglobulin light chains of the same type, ie, either immunoglobulin kappa (Igκ) or lambda (Igλ) light chains. Cell surface IgM serves as a receptor that can specifically bind to antigens, including those expressed by microbial pathogens, and trigger the clonal expansion and differentiations of B cells into antibody-secreting plasma cells. A soluble pentameric form of IgM can be produced and secreted by plasma cells into the blood. Pentameric IgM is comprised of 5 monomers that consist of two Igµ heavy chains and two light chains that are complexed with a polypeptide J-chain. The UCH-B1 monoclonal antibody recognizes both cell surface and soluble human IgM. IgM is an important component in the first line of defense against foreign pathogens, but may also play a role in autoimmune diseases. The UCH-B1 antibody can reportedly activate or costimulate the proliferation of normal B cells or some transformed B cell lines.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific BlueTM conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.
Development References (4)
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Armitage RJ, Rowe DJ, Beverly PC. A new antigen identified by the monoclonal antibody UCHB 1 delivers a costimulatory signal to a subset of human B cells.. Eur J Immunol. 1988; 18(1):67-76. (Immunogen: Activation, Calcium Flux, (Co)-stimulation, Flow cytometry, Functional assay). View Reference
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Bain B, Morilla R, Monard S, Kokai Y, Catovsky D. Spectrum of Reactivity with Three Monoclonal Antibodies-MHM6(CD23), L30(CD24) and UCHB1-in B-Cell Leukaemias.. Leuk Lymphoma. 1990; 3(2):97-102. (Clone-specific: Immunocytochemistry, Immunofluorescence). View Reference
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Klymenko T, Bloehdorn J, Bahlo J, et al. Lamin B1 regulates somatic mutations and progression of B-cell malignancies.. Leukemia. 2018; 32(2):364-375. (Clone-specific: Cell separation). View Reference
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Smith-Ravin J, Spencer J, Beverley PC, Isaacson PG. Characterization of two monoclonal antibodies (UCL4D12 and UCL3D3) that discriminate between human mantle zone and marginal zone B cells.. Clin Exp Immunol. 1990; 82(1):181-7. (Clone-specific: ELISA, Flow cytometry). 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.