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Multiparameter flow cytometric analysis of CD34 expression on Human CD14-negative peripheral blood mononuclear cells (PBMC). Human PBMC were stained with PE Mouse Anti-Human CD14 antibody (Cat. No. 555398/557154/561707) and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat No. 562438, Left Plot) or BD Horizon™ BV421 Mouse Anti-Human CD34 antibody (Cat No. 568758, Right Plot) at 1.0 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD34 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) CD14-negative PBMC. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ BV421 Mouse Anti-Human CD34
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information is found in the Technical Data Sheet for BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Companion Products
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.
Development References (14)
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Andrews RG, Singer JW, Bernstein ID. Precursors of colony-forming cells in humans can be distinguished from colony-forming cells by expression of the CD33 and CD34 antigens and light scatter properties.. J Exp Med. 1989; 169(5):1721-31. (Biology). View Reference
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Brocklebank AM, Sparrow RL. Enumeration of CD34+ cells in cord blood: a variation on a single-platform flow cytometric method based on the ISHAGE gating strategy.. Cytometry. 2001; 46(4):254-61. (Biology). View Reference
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Gore SD, Kastan MB, Civin CI. Normal human bone marrow precursors that express terminal deoxynucleotidyl transferase include T-cell precursors and possible lymphoid stem cells.. Blood. 1991; 77(8):1681-90. (Biology). View Reference
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Greaves MF, Titley I, Colman SM, et al. CD34 cluster workshop report. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:840-846.
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Hurwitz CA, Loken MR, Graham ML, et al. Asynchronous antigen expression in B lineage acute lymphoblastic leukemia.. Blood. 1988; 72(1):299-307. (Biology). View Reference
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Kurtzberg J, Denning SM, Nycum LM, Singer KH, Haynes BF. Immature human thymocytes can be driven to differentiate into nonlymphoid lineages by cytokines from thymic epithelial cells.. Proc Natl Acad Sci USA. 1989; 86(19):7575-9. (Biology). View Reference
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Lansdorp PM, Sutherland HJ, Eaves CJ. Selective expression of CD45 isoforms on functional subpopulations of CD34+ hemopoietic cells from human bone marrow.. J Exp Med. 1990; 172(1):363-6. (Clone-specific: Flow cytometry). View Reference
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Lanza F, Moretti S, Papa S, Malavasi F, Castoldi G. Report on the Fifth International Workshop on Human Leukocyte Differentiation Antigens, Boston, November 3-7, 1993.. Haematologica. 79(4):374-86. (Clone-specific: Flow cytometry). View Reference
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Leary AG, Strauss LC, Civin CI, Ogawa M. Disparate differentiation in hemopoietic colonies derived from human paired progenitors.. Blood. 1985; 66(2):327-32. (Biology). View Reference
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Loken MR, Shah VO, Dattilio KL, Civin CI. Flow cytometric analysis of human bone marrow. II. Normal B lymphocyte development. Blood. 1987; 70(5):1316-1324. (Biology). View Reference
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Loken MR, Shah VO, Dattilio KL, Civin CI. Flow cytometric analysis of human bone marrow: I. Normal erythroid development.. Blood. 1987; 69(1):255-63. (Biology). View Reference
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Ryan D, Kossover S, Mitchell S, Frantz C, Hennessy L, Cohen H. Subpopulations of common acute lymphoblastic leukemia antigen-positive lymphoid cells in normal bone marrow identified by hematopoietic differentiation antigens.. Blood. 1986; 68(2):417-25. (Biology). View Reference
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Siena S, Bregni M, Brando B, et al. Flow cytometry for clinical estimation of circulating hematopoietic progenitors for autologous transplantation in cancer patients.. Blood. 1991; 77(2):400-9. (Clone-specific: Flow cytometry). View Reference
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Terstappen LW, Huang S, Safford M, Lansdorp PM, Loken MR. Sequential generations of hematopoietic colonies derived from single nonlineage-committed CD34+CD38- progenitor cells. Blood. 1991; 77(6):1218-1227. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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