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BV421 Rat Anti-Mouse CD34
BV421 Rat Anti-Mouse CD34
Multicolor flow cytometric analysis of CD34 expression on mouse bone marrow cells.   BALB/c bone marrow cells were treated with Mouse BD Fc Block™ [Purified Rat Anti-Mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/ 553142)] and stained with either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Left Panel) or BD Horizon™ BV421 Rat Anti-Mouse CD34 antibody (Right Panel) followed by staining with APC Mouse Lineage Antibody Cocktail (Cat. No. 558074) to identify major lineage-committed cell types. Two-color flow cytometric contour plots were derived from gated events with the forward and side light-scatter characteristics of viable bone marrow leukocytes. Flow cytometry was performed using a BD FACSCanto™ II Flow Cytometer System.
Multicolor flow cytometric analysis of CD34 expression on mouse bone marrow cells.   BALB/c bone marrow cells were treated with Mouse BD Fc Block™ [Purified Rat Anti-Mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/ 553142)] and stained with either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Left Panel) or BD Horizon™ BV421 Rat Anti-Mouse CD34 antibody (Right Panel) followed by staining with APC Mouse Lineage Antibody Cocktail (Cat. No. 558074) to identify major lineage-committed cell types. Two-color flow cytometric contour plots were derived from gated events with the forward and side light-scatter characteristics of viable bone marrow leukocytes. Flow cytometry was performed using a BD FACSCanto™ II Flow Cytometer System.
Product Details
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BD Horizon™
Cd34; CD34 antigen; Hematopoietic progenitor cell antigen CD34
Mouse (QC Testing)
Rat IgG2a, κ
Recombinant Mouse CD34
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_11154576
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


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 the dye under optimum conditions and unconjugated antibody and free dye were removed.

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 can be found in the Technical Data Sheet of the BD Horizon Brilliant™ Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant™ Stain Buffer Plus (Cat. No. 566385).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  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/us/s/resources for technical protocols.
  8. 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.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
562608 Rev. 3
Antibody Details
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RAM34

The RAM34 monoclonal antibody specifically binds to the CD34 glycoprotein on the surface of three independently derived mouse CD34-transfected cell lines. RAM34 antibody also reacts with the mouse cell lines PA6, 416B, Swiss 3T6, NIH, 3T3, DA1, and M1, all of which are positive for expression of mouse CD34 mRNA. Cell lines shown to be negative for CD34 mRNA transcript, including WEHI-3B, EL4, 18.8, and CMT64/61, are also negative for surface expression of CD34 as determined by RAM34 staining. Normal thymocytes and splenocytes are negative for CD34 expression. In the bone marrow, 7-10% of cells are stained with the RAM34 antibody, including most of the Ly-6A/E (Sca-1)+ CD90 (Thy-1)low Lineage Marker- hematopoietic stem cell-enriched subpopulation and myeloerythroid progenitors. CD34 is also expressed on a small percentage of fetal liver cells, including NK-cell progenitors. CD34 has been reported to be expressed on the endothelium of capillaries and, in this form, to function as a ligand for L-selectin. Consistent with this observation, RAM34 antibody stains endothelial cells in spleen, thymus, and postcapillary HEVs in the lymph nodes. It is reported that RAM34 antibody can be used to select CD34+ CD117 (c-Kit)+ Ly-6A/E (Sca-1)+ Lineage Marker- bone marrow-derived hematopoietic stem cells, capable of short-term multi-lineage reconstitution of lethally irradiated mice. In  contrast, the  CD34- CD117+ Sca-1+ Lineage Marker- population contains self-renewing hematopoietic stem cells. Similarly, the bone marrow population with high dye-efflux capacity and highly enriched for long-term reconstituting hematopoietic stem cells, is CD34- CD117 (c-Kit)+ Ly-6A/E (Sca-1)+ Lineage Marker-.

        The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon™ BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon™ BV421, Pacific Blue™, and BD Horizon™ V450 cannot be used simultaneously.

562608 Rev. 3
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
562608 Rev.3
Citations & References
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Development References (11)

  1. Akashi K, Traver D, Miyamoto T, Weissman IL. A clonogenic common myeloid progenitor that gives rise to all myeloid lineages. Nature. 2000; 404(6774):193-197. (Clone-specific: Flow cytometry). View Reference
  2. Baumheter S, Singer MS, Henzel W, et al. Binding of L-selectin to the vascular sialomucin CD34. Science. 1993; 262(5132):436-438. (Biology). View Reference
  3. Brown J, Greaves MF, Molgaard HV. The gene encoding the stem cell antigen, CD34, is conserved in mouse and expressed in haemopoietic progenitor cell lines, brain, and embryonic fibroblasts. Int Immunol. 1991; 3(2):175-184. (Biology). View Reference
  4. Goodell MA, Rosenzweig M, Kim H, et al. Dye efflux studies suggest that hematopoietic stem cells expressing low or undetectable levels of CD34 antigen exist in multiple species. Nat Med. 1997; 3(12):1337-1345. (Clone-specific: Flow cytometry). View Reference
  5. Lorenz K, Grashoff C, Torka R, et al. Integrin-linked kinase is required for epidermal and hair follicle morphogenesis. J Cell Biol. 2007; 177(3):501-513. (Biology: Immunofluorescence, Immunohistochemistry). View Reference
  6. Lu J, Patrene KD, Herberman RB, Boggs SS. Expression of murine CD34 by fetal liver NK cell progenitors. Exp Hematol. 1999; 27(2):272-281. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  7. Morel F, Szilvassy SJ, Travis M, Chen B, Galy A. Primitive hematopoietic cells in murine bone marrow express the CD34 antigen. Blood. 1996; 88(10):3774-3784. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  8. Osawa M, Hanada K, Hamada H, Nakauchi H. Long-term lymphohematopoietic reconstitution by a single CD34-low/negative hematopoietic stem cell. Science. 1996; 273(5272):242-245. (Immunogen: Flow cytometry). View Reference
  9. Spangrude GJ, Heimfeld S, Weissman IL. Purification and characterization of mouse hematopoietic stem cells. Science. 1988; 241(4861):58-62. (Biology). View Reference
  10. Suda J, Sudo T, Ito M, Ohno N, Yamaguchi Y, Suda T. Two types of murine CD34 mRNA generated by alternative splicing. Blood. 1992; 79(9):2288-2295. (Biology). View Reference
  11. Suzuki A, Andrew DP, Gonzalo JA, et al. CD34-deficient mice have reduced eosinophil accumulation after allergen exposure and show a novel crossreactive 90-kD protein. Blood. 1996; 87(9):3550-3562. (Clone-specific: Immunoaffinity chromatography, Immunohistochemistry, Immunoprecipitation). View Reference
View All (11) View Less
562608 Rev. 3

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.