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).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- BD Horizon Brilliant Violet 650 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 7E9 antibody specifically recognizes CD21/CD35, an epitope shared by mouse Complement receptor 2 (CR2) and Complement receptor 1 (CR1), which are also known as CD21 and CD35, respectively. Mouse CD21 and CD35 are ~145-kDa and ~190-kDa single pass type I transmembrane glycoproteins, respectively. Unlike human CD21 and CD35 that are encoded by separate CR2 and CR1 genes, mouse CD21 and CD35 are encoded by a single Cr2 gene and are generated by alternative mRNA splicing. CD21 and CD35 are expressed on B cells and follicular dendritic cells (FCD) but not on thymocytes, T cells, erythrocytes, or platelets. CD35 is also expressed on macrophages and activated granulocytes. CD21 binds to cleaved C3b fragments, including C3d- and C3dg-bound to antigen, whereas CD35 binds to C3b-, iC3b-, and C4b-bound complexes. These receptors play many important roles including the transport, presentation, and retention of complement fragment-tagged antigens and immune complexes crucial for generating strong B cell antibody-producing responses. Complement-tagged antigen can bind to CD21, part of the B cell coreceptor complex and to the B cell receptor for antigen (BCR) which facilitates the antigen-driven activation of B cell responses. CD35 interacts with other regulatory factors to promote C3b and C4b degradation as well as playing roles in stimulating phagocytic functions in activated leucocytes. The 7E9 antibody recognizes an epitope on CD35 distinct from the epitope recognized by the mouse CD35-specific clone, 8C12, as well as the mouse CD21/CD35-specific clone, 7G6.
The antibody was conjugated to BD Horizon™ BV650 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm. BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor™ 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
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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.