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APC Rat Anti-Mouse CD304
APC Rat Anti-Mouse CD304

Multicolor flow cytometric analysis of CD304 expression on mouse splenic lymphocytes. Balb/c mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were stained with PE Rat Anti-Mouse CD4 (Cat. No. 561829/553730/557308), and APC Rat IgG2a, κ Isotype Control (Cat. No. 553932; Left Plot) or APC Rat Anti-Mouse CD304 antibody (Cat. No. 567461; 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 preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were stained with PE Rat Anti-Mouse CD4 (Cat. No. 561829/553730/557308), and APC Rat IgG2a, κ Isotype Control (Cat. No. 553932; Left Plot) or APC Rat Anti-Mouse CD304 antibody (Cat. No. 567461; 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 Pharmingen™
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 ≤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 to ensure that BD® CompBeads are appropriate for your specific cellular application.

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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
567461 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.

567461 Rev. 1
Format Details
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC
Red 627-640 nm
651 nm
660 nm
567461 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
567461 Rev. 1

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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.