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).
For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- 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.
- The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
- 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 refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The U21-1283 monoclonal antibody specifically recognizes Neuropilin-1 (NRP1), also known as CD304, Blood dendritic cell antigen 4 (BDCA4), and Vascular endothelial cell growth factor 165 receptor (VEGF165R). CD304 is a type I transmembrane glycoprotein involved in the development of the nervous and cardiovascular systems. It mediates the interaction, growth, survival, and migration of a variety of normal and tumor cells. CD304 is expressed on neurons, thymocytes, regulatory T cells, a subset of T follicular helper cells, dendritic cells, endothelial cells, and certain tumor cells. Neuropilin-1 has a very short cytoplasmic domain and interacts with various coreceptors to form ligand-binding, signal-transducing receptor complexes. CD304 complexes with Plexin-A family members to bind chemorepellent Class 3 Semaphorins and guide neuronal axon growth. It also functions as a coreceptor with VEGFR2/CD309 to stimulate angiogenesis in response to VEGF165. CD304 mediates the interactions between some T cells and dendritic cells.
The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.
Development References (6)
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Bruder D et al. Neuropilin-1: a surface marker of regulatory T cells.. Eur J Immunol. 2004; 34(3):623-630. (Biology). View Reference
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Dzionek A et al. BDCA-2, BDCA-3, and BDCA-4: three markers for distinct subsets of dendritic cells in human peripheral blood.. J Immunol. 2000; 165(11):6037-6046. (Biology). View Reference
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Mizui M, Kumanogoh A, Kikutani H. Immune semaphorins: novel features of neural guidance molecules.. J Clin Immunol. 2009; 29(1):1-11. (Biology). View Reference
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Pan Q et al. Neuropilin-1 binds to VEGF121 and regulates endothelial cell migration and sprouting.. J Biol Chem. 2007; 282(33):24049-24056. (Biology). View Reference
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Renand A, Milpied P, Rossignol J, et al. Neuropilin-1 expression characterizes T follicular helper (Tfh) cells activated during B cell differentiation in human secondary lymphoid organs.. PLoS ONE. 2013; 8(12):e85589. (Biology). View Reference
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Soker S, Takashima S, Miao HQ, Neufeld G, Klagsbrun M. Neuropilin-1 is expressed by endothelial and tumor cells as an isoform-specific receptor for vascular endothelial growth factor.. Cell. 1998; 92(6):735-45. (Biology). View Reference
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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.