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 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® CompBeads 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 BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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 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.
- 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).
Companion Products
The ZAQ2 monoclonal antibody specifically recognizes CD300LG, a microvascular endothelial adhesion molecule that belongs to the CD300 antigen like family. CD300LG is also known as Nepmucin (a not expressed in Peyer's patches mucin) and CMRF35-like molecule 9 (CLM-9). Four isoforms of this type I transmembrane glycoprotein have been described that are encoded by Cd300lg. The extracellular region of this sialomucin contains one IgV-like domain and one mucin-like domain followed by a transmembrane region and an intracellular tail. The ZAQ2 antibody recognizes the Ig domain of CD300LG (Nepmucin). This sialomucin is variably expressed on the luminal surfaces of endothelial cells lining small arterioles, venules and capillaries in many tissues. Although CD300LG (Nepmucin) protein is expressed by high endothelial venules (HEV) in lymph nodes, it is not expressed by Peyer's patches HEV. Nepmucin mRNA expression has also been observed for some myeloid lineage cell lines as well as CD11c+ dendritic cells, but not purified T and B lymphocytes. HEV-associated CD300LG (Nepmucin) mediates homotypic interactions between endothelial cells. This sialomucin also supports L-selectin-dependent lymphocyte rolling and heterophilic binding of lymphocytes by endothelial cells leading to transendothelial migration (TEM) of lymphocytes. The ZAQ2 antibody can reportedly inhibit lymphocyte TEM. CD300LG (Nepmucin) expression on microvascular endothelial cells is affected by local factors generated in tissues, including sites undergoing inflammation or tumor cell infiltration. This may in turn affect the levels of lymphocytic infiltration influenced by this sialomucin.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
Development References (4)
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Jin S, Umemoto E, Tanaka T, et al. Nepmucin/CLM-9, an Ig domain-containing sialomucin in vascular endothelial cells, promotes lymphocyte transendothelial migration in vitro.. FEBS Lett. 2008; 582(20):3018-24. (Clone-specific: Functional assay, Immunofluorescence, Inhibition). View Reference
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Umemoto E, Hayasaka H, Bai Z, et al. Novel regulators of lymphocyte trafficking across high endothelial venules.. Crit Rev Immunol. 2011; 31(2):147-69. (Biology). View Reference
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Umemoto E, Takeda A, Jin S, et al. Dynamic changes in endothelial cell adhesion molecule nepmucin/CD300LG expression under physiological and pathological conditions.. PLoS ONE. 2013; 8(12):e83681. (Clone-specific: Immunofluorescence, Immunohistochemistry). View Reference
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Umemoto E, Tanaka T, Kanda H, et al. Nepmucin, a novel HEV sialomucin, mediates L-selectin-dependent lymphocyte rolling and promotes lymphocyte adhesion under flow.. J Exp Med. 2006; 203(6):1603-14. (Immunogen: ELISA, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western blot). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.