Multicolor flow cytometric analysis of CD304 expression on Mouse splenic lymphocytes. BALB/c Mouse splenocytes were stained with BD Horizon™ BUV395 Rat Anti-Mouse CD4 antibody (Cat. No. 563790/565974), and with either APC-Cy7 Rat IgG2a, κ Isotype Control (Cat No. 552770; Left Plot) or APC-Cy7 Rat Anti-Mouse CD304 antibody (Cat No. 567698; Right Plot) at 0.5 ug/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the 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 (DAPI-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer 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 BD Horizon™ BUV395 Rat Anti-Mouse CD4 antibody (Cat. No. 563790/565974), and with either APC-Cy7 Rat IgG2a, κ Isotype Control (Cat No. 552770; Left Plot) or APC-Cy7 Rat Anti-Mouse CD304 antibody (Cat No. 567698; Right Plot) at 0.5 ug/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the 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 (DAPI-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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
18186
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO
Preparation And Storage
The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- APC-Cy7 is a tandem fluorochrome composed of Allophycocyanin (APC), which is excited by laser lines between 595 and 647 nm and serves as an energy donor, coupled to the cyanine dye Cy7™, which acts as an energy acceptor and fluoresces at 780 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in APC-Cy7, thus maximizing its fluorescence emission intensity, minimizing residual emission from APC, and minimizing required electronic compensation in multilaser-laser flow cytometry systems. Note: Although every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-Cy7 conjugate.
- APC-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
APC-Cy™7 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
552770
BUV395 Rat Anti-Mouse CD4 RUO
Size
50 µg
Cat No.
563790
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
DAPI Solution RUO
Size
1 mg
Cat No.
564907
567698 Rev. 2
Antibody Details
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.
567698 Rev. 2
Format Details
APC-Cy7
APC-Cy7 dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of a Allophycocyanin (APC) donor that has excitation maxima (Ex Max) of 651 nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 779 nm. APC-Cy7 can be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 780 nm (e.g., a 760/60 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
APC-Cy7
Red 627-640 nm
651 nm
779 nm
567698 Rev.2
Citations & References
View product citations for antibody "567698" on CiteAb
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: Flow cytometry). View Reference
567698 Rev. 2
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.