BD Horizon™ Technical Data Sheet BV510 Mouse Anti-Human CD238 Product Information Material Number: Alternate Name: Size: Vol. per Test: Clone: Immunogen: Isotype: Reactivity: Workshop: Storage Buffer: 563475 KEL; KELL; Kell blood group, metallo-endopeptidase; ECE3 50 tests 5 µl BRIC 203 Human erythrocytes Mouse IgG1, κ QC Testing: Human VII 70565 Aqueous buffered solution containing BSA and ≤0.09% sodium azide. Description The BRIC 203 monoclonal antibody specifically binds to the Kell blood group antigen, Kp(a-b+) but not Kp(a+b-), also known as the Kell blood group metallo-endopeptidase, KELL, ECE3 and CD238. CD238 is an approximately 115-kDa type II transmembrane glycoprotein that is encoded by the KEL gene. CD238 functions as a zinc endopeptidase that cleaves endothelin-3 into an active form which can function as a vasoconstrictor. CD238 is expressed on erythrocytes, human embryonic stem cell-derived endoderm and on cells from various other tissues, including testis, heart, brain and skeletal muscle. Decreased expression of CD238 is associated with the McLeod phenotype as a consequence of losing XK protein expression. Kell antigen expression can play an important role during blood transfusions and also specific autoimmune diseases related to Kell antigen recognition. During development the BRIC 203 monoclonal antibody was found to detect the CD238 antigen by flow cytometric analysis of human erythrocytes. It reportedly works in immunoprecipitation and agglutination assays. The antibody was conjugated to BD Horizon™ BV510 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 405-nm and Em Max at 510-nm, BD Horizon™ BV510 can be excited by the violet laser and detected in the BD Horizon™ V500 (525/50-nm) filter set. BD Horizon™ BV510 conjugates are useful for the detection of dim markers off the violet laser. Flow cytometric analysis of CD238 expression on human erythrocytes . Human erythrocytes were stained with the BD Horizon™ BV510 Mouse Anti-Human CD238 antibody (Cat. No. 563475; solid line histogram) or with BD Horizon™ BV510 Mouse IgG1, κ Isotype Control (Cat. No. 562946; dashed line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of erythrocytes. Flow cytometric analysis was performed using a BD™ LSR II 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™ BV510 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV510 were removed. Application Notes Application Flow cytometry 563475 Rev. 1 Routinely Tested Page 1 of 2 Suggested Companion Products Catalog Number 554656 562946 Name Stain Buffer (FBS) BV510 Mouse IgG1, k Isotype Control Size 500 ml 50 µg Clone (none) X40 Product Notices 1. 2. 3. 4. 5. 6. 7. 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). Source of all serum proteins is from USDA inspected abattoirs located in the United States. An isotype control should be used at the same concentration as the antibody of interest. Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols. 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. Brilliant Violet™ 510 is a trademark of Sirigen. References Jung HH, Hergersberg M, Vogt M, Pahnke J, Treyer V, Rothlisberger B, Kollias SS, Russo D, Frey BM. McLeod phenotype associated with a XK missense mutation without hematologic, neuromuscular, or cerebral involvement. Transfusion. 2003; 43(7):928-938. (Clone-specific: Flow cytometry) Parsons SF, Gardner B, Anstee DJ. Monoclonal antibodies against Kell glycoprotein: serology, immunochemistry and quantification of antigen sites. Transfus Med. 1993; 3(2):137-142. (Immunogen) van der Schoot CE, Baardman R, Lighthart P, de Jong I, von dem Borne AEK, de Haas M. CD238 - Kell. In: Mason D, Andre P, Bensussan A, et al, ed. Leucocyte Typing VII : White Cell Differentiation Antigens. New York: Oxford University Press; 2002:584-585. (Clone-specific) Wang P, Rodriguez RT, Wang J, Ghodasara A, Kim SK. Targeting SOX17 in Human Embryonic Stem Cells Creates Unique Strategies for Isolating and Analyzing Developing Endoderm. Cell Stem Cell. 2011; 8:335-346. (Biology) 563475 Rev. 1 Page 2 of 2