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BUV395 Rat Anti-Mouse CD304
BUV395 Rat Anti-Mouse CD304
Multicolor flow cytometric analysis of CD304 expression on mouse splenic lymphocytes. Balb/c mouse splenocytes were stained with PE Rat Anti-Mouse CD4 (Cat. No. 561829/553730/557308), and BD Horizon™ BUV395 Rat IgG2a, κ Isotype Control (Cat. No. 563556; Left Plot) or BD Horizon™ BUV395 Rat Anti-Mouse CD304 antibody (Cat. No. 567452; Right Plot) at 0.5 μg/test. 7-AAD (7-Amino-Actinomycin D) Solution (Cat. No. 559925) was added to the cells right before analysis. Bivariate pseudocolor density plots showing correlated expression of CD304 (or Ig Isotype control) versus CD4 staining was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD X-20 LSRFortessa™ Cell Analyze System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multicolor flow cytometric analysis of CD304 expression on mouse splenic lymphocytes. Balb/c mouse splenocytes were stained with PE Rat Anti-Mouse CD4 (Cat. No. 561829/553730/557308), and BD Horizon™ BUV395 Rat IgG2a, κ Isotype Control (Cat. No. 563556; Left Plot) or BD Horizon™ BUV395 Rat Anti-Mouse CD304 antibody (Cat. No. 567452; Right Plot) at 0.5 μg/test. 7-AAD (7-Amino-Actinomycin D) Solution (Cat. No. 559925) was added to the cells right before analysis. Bivariate pseudocolor density plots showing correlated expression of CD304 (or Ig Isotype control) versus CD4 staining was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD X-20 LSRFortessa™ Cell Analyze System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Horizon™
neuropilin-1; Nrp1; A5 protein; NP-1; NPN-1; Npn1; Nrp
Mouse (QC Testing)
Rat WI, also known as Wistar (outbred) IgG2a, κ
Mouse CD304 Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.

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. This will 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

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
  8. 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.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
567452 Rev. 1
Antibody Details
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V46-1954

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.

The antibody was conjugated to BD Horizon™ BUV395 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. With an Ex Max near 348 nm and an Em Max near 395 nm, BD Horizon™ BUV395 can be excited by the ultraviolet laser (355 nm) laser and detected with a 379/28 filter. This dye has been exclusively developed by BD Biosciences as an optimal dye for use on instruments equipped with the ultraviolet laser and has virtually no spillover into any other detector.

567452 Rev. 1
Format Details
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BUV395
The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV395
Ultraviolet 355 nm
348 nm
395 nm
567452 Rev.1
Citations & References
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Development References (9)

  1. 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
  2. Goel HL, Mercurio AM. VEGF targets the tumour cell.. Nat Rev Cancer. 2013; 13(12):871-82. (Biology). View Reference
  3. Guo HF, Vander Kooi CW. Neuropilin Functions as an Essential Cell Surface Receptor.. J Biol Chem. 2015; 290(49):29120-6. (Biology). View Reference
  4. 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
  5. 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
  6. 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.
  7. 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
  8. 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
  9. 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
View All (9) View Less
567452 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.