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Alexa Fluor® 700 Rat Anti-Mouse CD34
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Alexa Fluor® 700 Rat Anti-Mouse CD34

Flow cytometric analysis for CD34 in mouse bone marrow. BALB/c bone marrow cells were treated with Mouse BD Fc Block™ (Purified Rat Anti-Mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141)) and stained with either the Alexa Fluor® 700 Rat Anti-Mouse CD34 antibody (left and right panels) or a Alexa Fluor® 700 Rat IgG2a, κ isotype control (left and middle panels), followed by staining with an APC Lineage Cocktail (Cat. No. 558074) to identify major lineage committed cells. Histogram and contour plots were derived from the gated events based on the light scattering characteristics of leukocytes and fluorescence characteristics of the CD34 and Lineage markers. Flow cytometry was performed on a BD LSR™ II flow cytometry system.

Flow cytometric analysis for CD34 in mouse bone marrow. BALB/c bone marrow cells were treated with Mouse BD Fc Block™ (Purified Rat Anti-Mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141)) and stained with either the Alexa Fluor® 700 Rat Anti-Mouse CD34 antibody (left and right panels) or a Alexa Fluor® 700 Rat IgG2a, κ isotype control (left and middle panels), followed by staining with an APC Lineage Cocktail (Cat. No. 558074) to identify major lineage committed cells. Histogram and contour plots were derived from the gated events based on the light scattering characteristics of leukocytes and fluorescence characteristics of the CD34 and Lineage markers. Flow cytometry was performed on a BD LSR™ II flow cytometry system.

Product Details
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BD Pharmingen™
Mouse (QC Testing)
Rat IgG2a, κ
Recombinant Mouse CD34
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_1727471
Aqueous buffered solution containing protein stabilizer 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 Alexa Fluor® 700 under optimum conditions, and unreacted Alexa Fluor® 700 was removed.

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. Alexa Fluor® 700 has an adsorption maximum of ~700nm and a peak fluorescence emission of ~720nm. Before staining cells with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  5. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
560518 Rev. 2
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-.

560518 Rev. 2
Format Details
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Alexa Fluor™ 700
Alexa Fluor™ 700 dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 697 nm and an emission maximum (Em Max) at 719-nm. Alexa Fluor™ 700 is designed to be excited by the Red (627–640-nm) laser and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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Alexa Fluor™ 700
Red 627-640 nm
697 nm
719 nm
560518 Rev.2
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
560518 Rev. 2

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

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