Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) 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).
The 8G12 monoclonal antibody specifically recognizes CD34, a 105-120 kDa single-chain type I transmembrane glycoprotein. The 8G12 antibody recognizes an epitope on CD34 distinct from the one recognized by clone My10. CD34 is expressed on immature hematopoietic precursor cells and all hematopoietic colony-forming cells in bone marrow and blood, including unipotent (CFU-GM, BFU-E) and pluripotent progenitors (CFU-GEMM, CFU-Mix, and CFUBlast). The CD34 antigen is a differentiation stage-specific leucocyte antigen. Terminal deoxynucleotidyl transferase-positive B- and T-lymphoid precursors in normal bone marrow are CD34+. The CD34 antigen is present on early myeloid cells that express the CD33 antigen but lack the CD14 and CD15 antigens and on early erythroid cells that express the CD71 antigen and dimly express the CD45 antigen. The CD34 antigen is also found on capillary endothelial cells and approximately 1% of human thymocytes. Normal peripheral blood lymphocytes, monocytes, granulocytes, and platelets do not express CD34. CD34 density is highest on early hematopoietic progenitor cells and decreases as cells mature. The antigen is absent on fully differentiated hematopoietic cells. Uncommitted CD34+ progenitor cells are CD38- and lack lineage-specific antigens such as CD71, CD33, CD10, and CD5, while CD34+ cells that are lineage-committed express the CD38 antigen in high density. Most CD34+ cells reciprocally express either the CD45RO or CD45RA antigens, with the CD45RO+ population being the more primitive. Approximately 60% of acute B-lymphoid leukemias and acute myeloid leukemias (AML) and 1% to 5% of acute T-lymphoid leukemias express CD34. CD34 is not expressed on chronic lymphoid leukemias or lymphomas.
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.
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