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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.
Product Notices
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products
The V46-1954 monoclonal antibody specifically recognizes mouse CD304, also known as neuropilin 1 (Nrp1). It does not cross-react with neuropilin 2 (Nrp2). CD304 is a type-1 transmembrane protein that plays important roles in neuronal development, angiogenesis, and immunosuppression. Through interactions with plexins and VEGF receptors on neurons and endothelial cells, it is a co-receptor for the class III semaphorin subfamily and vascular endothelial growth factor (VEGF), respectively. The ability of the S1 protein of SARS-CoV-2 virus to bind to Nrp1, a potential mechanism for cellular infection, is being investigated. In the immune system, CD304 is expressed on natural (thymus-derived) Treg cells and may be responsible for the unsuccessful immune regulation of tumor growth.
Development References (9)
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Delgoffe GM, Woo SR, Turnis ME, et al. Stability and function of regulatory T cells is maintained by a neuropilin-1-semaphorin-4a axis.. Nature. 2013; 501(7466):252-6. (Biology). View Reference
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Goel HL, Mercurio AM. VEGF targets the tumour cell.. Nat Rev Cancer. 2013; 13(12):871-82. (Biology). View Reference
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Guo HF, Vander Kooi CW. Neuropilin Functions as an Essential Cell Surface Receptor.. J Biol Chem. 2015; 290(49):29120-6. (Biology). View Reference
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Hansen W, Hutzler M, Abel S, et al. Neuropilin 1 deficiency on CD4+Foxp3+ regulatory T cells impairs mouse melanoma growth.. J Exp Med. 2012; 209(11):2001-16. (Biology). View Reference
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Hwang JY, Sun Y, Carroll CR, Usherwood EJ. Neuropilin-1 Regulates the Secondary CD8 T Cell Response to Virus Infection. mSphere. 2019; 4(3):e00221-19. (Biology). View Reference
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James L. Daly, Boris Simonetti, Carlos Antón-Págaro, et al. Neuropilin-1 is a host factor for SARS-CoV-2 infection. 2020. Available: https://doi.org/10.1101/2020.06.05.134114 2020, June 6.
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Liu WQ, Lepelletier Y, Montès M, et al. NRPa-308, a new neuropilin-1 antagonist, exerts in vitro anti-angiogenic and anti-proliferative effects and in vivo anti-cancer effects in a mouse xenograft model.. Cancer Lett. 2018; 414:88-98. (Biology). View Reference
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Roy S, Bag AK, Singh RK, Talmadge JE, Batra SK, Datta K. Multifaceted Role of Neuropilins in the Immune System: Potential Targets for Immunotherapy.. Front Immunol. 2017; 8:1228. (Biology). View Reference
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Yadav M, Louvet C, Davini D, et al. Neuropilin-1 distinguishes natural and inducible regulatory T cells among regulatory T cell subsets in vivo.. J Exp Med. 2012; 209(10):1713-22, S1-19. (Biology). 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.