Multiparameter flow cytometric analysis of CD319 (CRACC) expression on human peripheral blood leucocytes. Human whole blood was stained with BD Horizon™ BUV395 Mouse Anti-Human CD3 antibody (Cat. No. 564000/555899) and either PE Mouse IgG2b, κ Isotype Control (Cat. No. 555058; Left Plots) or PE Mouse Anti-Human CD319 (CRACC) antibody (Cat. No. 566840; Right Plots). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Bivariate pseudocolor density plots showing the correlated expression of CD319 (CRACC) [or Ig Isotype control staining] versus side light-scatter signals were derived from gated events with the forward and side light scatter characteristics of viable leucocyte populations (Top Plots). The bivariate plots showing the correlated expression of CD319 (CRACC) [or Ig Isotype control staining] versus CD3 were derived from gated events with the forward and side light scatter characteristics of viable lymphocytes (Bottom Plots). Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
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Multiparameter flow cytometric analysis of CD319 (CRACC) expression on human peripheral blood leucocytes. Human whole blood was stained with BD Horizon™ BUV395 Mouse Anti-Human CD3 antibody (Cat. No. 564000/555899) and either PE Mouse IgG2b, κ Isotype Control (Cat. No. 555058; Left Plots) or PE Mouse Anti-Human CD319 (CRACC) antibody (Cat. No. 566840; Right Plots). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Bivariate pseudocolor density plots showing the correlated expression of CD319 (CRACC) [or Ig Isotype control staining] versus side light-scatter signals were derived from gated events with the forward and side light scatter characteristics of viable leucocyte populations (Top Plots). The bivariate plots showing the correlated expression of CD319 (CRACC) [or Ig Isotype control staining] versus CD3 were derived from gated events with the forward and side light scatter characteristics of viable lymphocytes (Bottom Plots). Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Product Details
BD Pharmingen™
19A; 19A24 protein; CD319; CRACC; CS1; SLAMF7
Human (QC Testing)
Mouse BALB/c IgG2b, κ
Human extracellular CRACC
Flow cytometry (Routinely Tested)
5 µl/test
VIII 80452
AB_2869897
Aqueous buffered solution containing BSA, protein stabilizer, 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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
PE Mouse IgG2b, κ Isotype Control RUO
Size
0.1 mg
Cat No.
555058
BUV395 Mouse Anti-Human CD3 RUO
Size
25 Tests
Cat No.
564000
566840 Rev. 2
Antibody Details
162/CRACC
The 162 monoclonal antibody specifically binds to CRACC (CD2-like receptor activating cytotoxic cells) which is also known as CD319, CS1 (CD2 subset 1), 19A, or 19A24. CD319 (CRACC) is a ~66 kDa type I transmembrane glycoprotein that is encoded by SLAMF7 (SLAM family member 7) which belongs to the CD2 family within the Ig superfamily. CD319 (CRACC) is expressed on most NK cells and subsets of CD8+ T cells, CD4+ T cells, B cells, activated macrophages, and dendritic cells. CRACC reportedly plays roles in the activation, proliferation and effector functions of T cells, NK cells, and other leucocytes.
566840 Rev. 2
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
566840 Rev.2
Citations & References
View product citations for antibody "566840" on CiteAb
Development References (6)
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Boles KS, Mathew PA. Molecular cloning of CS1, a novel human natural killer cell receptor belonging to the CD2 subset of the immunoglobulin superfamily. Immunogenetics. 2001; 52(3-4):302-307. (Biology). View Reference
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Bouchon A, Cella M, Grierson HL, Cohen JI, Colonna M. Activation of NK cell-mediated cytotoxicity by a SAP-independent receptor of the CD2 family. J Immunol. 2001; 167(10):5517-5521. (Immunogen: Flow cytometry, Functional assay, Immunoprecipitation). View Reference
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Hagberg N, Theorell J, Schlums H, Eloranta ML, Bryceson YT, Ronnblom L. Systemic lupus erythematosus immune complexes increase the expression of SLAM family members CD319 (CRACC) and CD229 (LY-9) on plasmacytoid dendritic cells and CD319 on CD56(dim) NK cells. J Immunol. 2013; 191(6):2989-2998. (Clone-specific). View Reference
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Kim JR, Horton NC, Mathew SO, Mathew PA. CS1 (SLAMF7) inhibits production of proinflammatory cytokines by activated monocytes. Inflamm Res. 2013; 62(8):765-772. (Biology). View Reference
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Murphy JJ, Hobby P, Vilarino-Varela J, et al. A novel immunoglobulin superfamily receptor (19A) related to CD2 is expressed on activated lymphocytes and promotes homotypic B-cell adhesion. Biochem J. 2002; 361(Pt3):431-436. (Biology). View Reference
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Tassi I, Colonna M. The cytotoxicity receptor CRACC (CS-1) recruits EAT-2 and activates the PI3K and phospholipase Cgamma signaling pathways in human NK cells. J Immunol. 2005; 175(12):7996-8002. (Clone-specific: Flow cytometry, Functional assay). View Reference
566840 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.