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BD Pharmingen™ Technical Data Sheet APC Mouse Anti-Human Angiotensin Converting Enzyme (CD143) Product Information 557929 ACE1; DCP1; Kininase II 100 Tests 20 µl BB9 Human bone marrow-derived stromal cells Mouse IgG1, κ QC Testing: Human Aqueous buffered solution containing BSA and ≤0.09% sodium azide. Material Number: Alternate Name: Size: Vol. per Test: Clone: Immunogen: Isotype: Reactivity: Storage Buffer: Description The BB9 Monoclonal antibody specifically binds Angiotensin-converting enzyme (ACE) which is also known as Dipeptidyl carboxypeptidase 1 (DCP1) or CD143. CD143 is a type I transmembrane glycoprotein that functions as a zinc metallopeptidase involved with the metabolism of angiotensin, bradykinin, and other oligopeptides. It is expressed on primitive hematopoietic progenitor cells (HPC). BB9 reacts with stromal cells, a minor subset of human bone marrow cells; it does not react with peripheral blood leucocytes. BB9 recognizes a subset of CD34+CD90+ cells that demonstrate low or absent expression of CD38, low retention of Rhodamine 123 and are Ki-67-negative. CD34+BB9+ cells are able to sustain hematopoiesis in pre-CFU culture stimulated with IL-3, IL-6, G-CSF, and SCF. BB9 also reacts with a subset of CD34+ cells from mobilized peripheral blood. This antibody was generated using human bone marrow-derived stromal cells as immunogen. Flow cytometric analysis of CD143 expression on human peripheral blood lymphocytes. Whole blood was stained with either APC Mouse Anti-Human Angiotensin Converting Enzyme (CD143) (Cat. No. 557929; solid line histogram) or APC Mouse IgG1, κ Isotype Control (Cat. No. 555751; dashed line histogram). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. 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 APC under optimum conditions, and unconjugated antibody and free APC were removed. Application Notes Application Flow cytometry Routinely Tested Suggested Companion Products Catalog Number 555751 554656 555899 557929 Rev. 5 Name APC Mouse IgG1, κ Isotype Control Stain Buffer (FBS) Lysing Buffer Size 100 Tests 500 mL 100 mL Clone MOPC-21 (none) (none) Page 1 of 2 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). Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser. 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. An isotype control should be used at the same concentration as the antibody of interest. References Jokubaitis VJ, Sinka L, Simmons PJ, et al. Angiotensin-converting enzyme (CD143) marks hematopoietic stem cells in human embryonic, fetal, and adult hematopoietic tissues. Blood. 2008; 111(8):4055-4063. (Clone-specific) Ramshaw HS, Haylock D, Swart B, et al. Monoclonal antibody BB9 raised against bone marrow stromal cells identifies a cell-surface glycoprotein expressed by primitive human hemopoietic progenitors. Exp Hematol. 2001; 29(8):981-992. (Immunogen) Sinka L, Biasch K, Khazaal I, Péault B, Tavian M. Angiotensin-converting enzyme (CD143) specifies emerging lympho-hematopoietic progenitors in the human embryo. Blood. 2012; 119(16):3712-3723. (Clone-specific) Zambidis ET, Park TS, Péault B, et al. Expression of angiotensin-converting enzyme (CD143) identifies and regulates primitive hemangioblasts derived from human pluripotent stem cells. Blood. 2008; 112(9):3601-3614. (Clone-specific) 557929 Rev. 5 Page 2 of 2