BD Horizon™ RB705 Mouse Anti-Human CD34
Clone 581 (RUO)
Multiparameter flow cytometric analysis of CD34 expression by viable Human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells were stained with FITC Mouse Anti-Human CD14 antibody (Cat. No. 555397) and with either BD Horizon™ RB705 Mouse IgG1, κ Isotype Control (Cat No. 570261; Left Plot) or BD Horizon™ RB705 Mouse Anti-Human CD34 antibody (Cat. No. 570578/570666; Right Plot). DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of side light-scatter (SSC-A) signals versus CD34 expression (or Ig Isotype control staining) was derived from CD14-negative gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
Multiparameter flow cytometric analysis of CD34 expression by viable Human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells were stained with FITC Mouse Anti-Human CD14 antibody (Cat. No. 555397) and with either BD Horizon™ RB705 Mouse IgG1, κ Isotype Control (Cat No. 570261; Left Plot) or BD Horizon™ RB705 Mouse Anti-Human CD34 antibody (Cat. No. 570578/570666; Right Plot). DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of side light-scatter (SSC-A) signals versus CD34 expression (or Ig Isotype control staining) was derived from CD14-negative gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
Product Details
BD Horizon™
gp105-120; My10; Hematopoietic progenitor cell antigen CD34
Human (QC Testing)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl/test
V MA27, VI E004
947
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
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 unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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.
Product Notices
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- 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.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
FITC Mouse Anti-Human CD14 RUO
Size
100 Tests
Cat No.
555397
RB705 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
570261
DAPI Solution RUO
Size
1 mg
Cat No.
564907
570666 Rev. 1
Antibody Details
581
The 581 monoclonal antibody specifically binds to CD34, a sialomucin-like type I transmembrane glycoprotein. This single-chain, 105-120 kDa, heavily O-glycosylated protein is expressed on hematopoietic progenitor cells, vascular endothelium, bone marrow stromal cells and embryonic fibroblasts. The cytoplasmic region of the CD34 antigen is a target for phosphorylation by activated protein kinase C suggesting CD34 may play a role in signal transduction. CD34 may also play a role as an adhesion molecule since it binds to CD62E and CD62L. Clone 581 binds to the class III CD34 epitope. It is resistant to neuraminidase, chymopapain and glycoprotease. The 581 antibody blocks reactivity of another anti-CD34 monoclonal antibody, 8G12.
570666 Rev. 1
Format Details
RB705
The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
RB705
Blue 488 nm
498 nm
707 nm
570666 Rev.1
Citations & References
View product citations for antibody "570666" on CiteAb
Development References (7)
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Cannavo N, Bossy D, Hirn J. CD34 Workshop: Increasing complexity of the CD34 epitope classification. In: Kishimoto T. Tadamitsu Kishimoto .. et al, ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:978-982.
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Egeland T, Tjonnfjord G, Steen R, Gaudernack G, Thorsby E. Positive selection of bone marrow-derived CD34 positive cells for possible stem cell transplantation. Transplant Proc. 1993; 25(1):1261-1263. (Biology). View Reference
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Gaudernack G, Egeland T. Epitope mapping of 33 CD34 mAb, including the Fith Workshop Panel. 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; 1995:861-864.
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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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Nishio H, Tada J, Hashiyama N, Hirn J, Ingles-Esteven J, Suda T. CD34. 1999. Available: http://mpr.nci.nih.gov/prow/guide/968267813_g.htm 2006, February 8.
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Owens MA, Loken MR. Peripheral blood stem cell quantitation. In: Owens MA, Loken MR. Flow Cytometry Principles for Clinical Laboratory Practice. New York: John Wiley & Sons; 1995:128.
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Zola H, Swart B, Nicholson I, Voss E. CD34. In: Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007:94.
570666 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
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.