BD Horizon™ Technical Data Sheet BV421 Mouse Anti-Human CD84 Product Information Material Number: 566094 Alternate Name: hCD84; SLAMF5; SLAF5; LY9B; Hly9-beta; Cell surface antigen MAX.3 Size: 50 Tests Vol. per Test: 5 µl Clone: 2G7 Immunogen: Namalwa Cell Line Isotype: Mouse IgG1, κ Reactivity: QC Testing: Human Workshop: V B057; VI B082 Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide. Description The 2G7 monoclonal antibody specifically binds to CD84, which is also known as SLAM family member 5 (SLAMF5), Hly9-beta, LY9B, or Cell surface antigen MAX.3. CD84 is 64-82 kDa type I transmembrane glycoprotein that belongs to the CD2 family within the Ig superfamily. CD84 is expressed on mature B cells, thymocytes, CD45RO+ T cells, NK cells, monocytes, and platelets. CD84 is also strongly expressed by tissue macrophages. CD84 may play a role as a homotypic adhesion molecule that mediates leucocyte interactions and signaling.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

Flow cytometric analysis of CD84 expression on human peripheral blood Two-color flow cytometric analysis of CD84 expression on human leucocytes. Whole blood was stained with either BD Horizon™ BV421 Mouse peripheral blood lymphocytes. Whole blood was stained with PE CD19 IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 (Cat. No. 555413/561741) and either BD Horizon™ BV421 Mouse IgG1, κ Mouse Anti-Human CD84 antibody (Cat. No. 566094; Right Plot). Erythrocytes Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon BV421 Mouse were lysed with BD FACS Lysing Solution (Cat. No. 349202). Flow cytometric Anti-Human CD84 antibody (Cat. No. 566094; Right Plot). Erythrocytes were plots showing the correlated expression of side scattered- signals (SSC) lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-parameter flow versus CD84 (or Ig Isotype Control staining) were derived from gated events cytometric plots showing the correlated expression of CD19 versus CD84 (or with the forward and side light-scatter characteristics of intact human Ig Isotype control staining) were derived from gated events with the forward peripheral blood leucocyte populations. Flow cytometric analysis was and side light-scatter characteristics of intact human peripheral blood performed using a BD LSRFortessa™ X-20 Flow Cytometer System. lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.

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 BD Horizon™ BV421 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV421 were removed. Application Notes Application Flow cytometry Routinely Tested

566094 Rev. 1 Page 1 of 2 Recommended Assay Procedure: For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

Suggested Companion Products Catalog Number Name Size Clone 554657 Stain Buffer (BSA) 500 mL (none) 554656 Stain Buffer (FBS) 500 mL (none) 562438 BV421 Mouse IgG1, k Isotype Control 50 µg X40 563794 Brilliant Stain Buffer 100 Tests (none) 555899 Lysing Buffer 100 mL (none) 349202 BD FACS™ Lysing Solution 100 mL (none) 555413 PE Mouse Anti-Human CD19 100 Tests HIB19 561741 PE Mouse Anti-Human CD19 25 Tests HIB19 Product Notices 1. 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). 2. An isotype control should be used at the same concentration as the antibody of interest. 3. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR. 4. Source of all serum is from USDA inspected abattoirs located in the United States. 5. 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. 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 www.bdbiosciences.com/pharmingen/protocols for technical protocols. References de la Fuente MA, Pizcueta P, Engel P. CD84 Workshop Panel report. In: Kishimoto T, Kikutani H, von dem Borne AEGK, et al, ed. Leucocyte Typing VI: White Cell Differentiation Antigens. New York, NY: Garland Publishing, Inc.; 1997:193-194. (Clone-specific: Flow cytometry) de la Fuente MA, Pizcueta P, Nadal M, Bosch J, Engel P. CD84 leukocyte antigen is a new member of the Ig superfamily. Blood. 1997; 90(6):2398-2405. (Clone-specific: Flow cytometry, Immunoprecipitation) Engel P, Smith H, Tedder TF. Phenotypic analysis with the B-cell Unknown Panel mAb. In: Schlossman SF, Boumsell L, Gilks W, et al, ed. Leukocyte Typing V: White Cell Differentiation Antigens. Oxford: Oxford University Press; 1995:607-620. (Clone-specific: Flow cytometry) Engel P, Wagner N, Tedder TF. CDw84 Workshop Report. In: Schlossman SF, Boumsell L, Gilks W, et al, ed. Leukocyte Typing V: White Cell Differentiation Antigens. Oxford: Oxford University Press; 1995:699-700. (Immunogen: Flow cytometry) Martin M, Romero X, de la Fuente MA, et al. CD84 functions as a homophilic adhesion molecule and enhances IFN-gamma secretion: adhesion is mediated by Ig-like domain 1. J Immunol. 2001; 167(7):3668-3676. (Biology) Romero X, Benítez D, March S, Vilella R, Miralpeix M, Engel P. Differential expression of SAP and EAT-2-binding leukocyte cell-surface molecules CD84, CD150 (SLAM), CD229 (Ly9) and CD244 (2B4). Tissue Antigens. 2004; 64(2):132-144. (Biology) Romero X, de la Fuente MA, Tovar V, et al. Characterization of a novel panel of CD84 monoclonal antibodies. In: Mason D, Andre P, Bensussan A, ed. Leucocyte Typing VII: White Cell Differentiation Antigens. New York: Oxford University Press; 2002:103-104. (Clone-specific: Immunoprecipitation) Sayós J, Martín M, Chen A, et al. Cell surface receptors Ly-9 and CD84 recruit the X-linked lymphoproliferative disease gene product SAP. Blood. 2001; 97(12):3867-3874. (Biology) Schlossman SF, Boumsell L, Gilks W, et al, ed. Leukocyte Typing V: White Cell Differentiation Antigens. Oxford: Oxford University Press; 1995(Clone-specific: Flow cytometry)

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