Skip to main content Skip to navigation
PE-Cy™5-Mouse Anti-Human CD34
PE-Cy™5-Mouse Anti-Human CD34
Multicolor flow cytometric analysis of CD34 expressed by human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells were incubated in the presence of Human BD Fc Block™(Cat. No. 564219/564220) and were stained with PE Mouse Anti-Human CD14 antibody (Cat. No. 555398/561707), FITC Mouse Anti-Human CD45 (Cat. No. 555482/560976/561865) and either a PE-Cy™5 Mouse IgG1 κ Isotype Control (Cat No. 555750, Left Panel) or the PE-Cy™5-Mouse Anti-Human CD34 antibody (Cat. No. 561819/555823, Right Panel). Flow cytometric dot plots showing side-scattered light signals versus CD34 fluorescence (or Ig isotype control staining) were derived from gated events based on the light scattering characteristics for viable CD14-negative cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of CD34 expressed by human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells were incubated in the presence of Human BD Fc Block™(Cat. No. 564219/564220) and were stained with PE Mouse Anti-Human CD14 antibody (Cat. No. 555398/561707), FITC Mouse Anti-Human CD45 (Cat. No. 555482/560976/561865) and either a PE-Cy™5 Mouse IgG1 κ Isotype Control (Cat No. 555750, Left Panel) or the PE-Cy™5-Mouse Anti-Human CD34 antibody (Cat. No. 561819/555823, Right Panel). Flow cytometric dot plots showing side-scattered light signals versus CD34 fluorescence (or Ig isotype control staining) were derived from gated events based on the light scattering characteristics for viable CD14-negative cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
Down Arrow Up Arrow


BD Pharmingen™
gp105-120; My10; Hematopoietic progenitor cell antigen CD34
Human (QC Testing)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
20 µl
V MA27, VI E004
947
AB_396152
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 with PE-Cy5 (formerly known as BD Cy-Chrome™) under optimum conditions, and unconjugated antibody and free PE-Cy5 were removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  7. PE-Cy5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the PE-Cy5 tandem fluorochrome, extra care must be taken when using dual-laser cytometers which may directly excite both PE and Cy5™.
  8. PE-Cy5 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by the 488 nm light of an Argon ion laser and serves as an energy donor, coupled to the cyanine dye Cy5, which acts as an energy acceptor and fluoresces at 670 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in PE-Cy5, thus maximizing its fluorescence emission intensity, minimizing residual emission from PE, and minimizing lot-to-lot variation.
  9. PE-Cy5 tandem fluorochromes have been reported to bind some classes of human macrophages and granulocytes via Fc receptors, and PE has been reported to bind to mouse B lymphocytes via Fc receptors. Preincubation of mouse leukocytes with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 can reduce the non-specific binding of PE-Cy5-conjugated reagents to mouse B cells. However, PE-Cy5 conjugated reagents should not be used to stain splenocytes of SJL, NOD, and MRL mice as B lymphocytes and/or other leukocytes have been reported to non-specifically stain regardless of the use of Mouse BD Fc Block™ (the CD72c complex has been implicated for PE-Cy5 binding in these strains). Reagents conjugated to PE, PerCP, PerCP-Cy5.5, APC, and APC-Cy7 tandem fluorochrome can be used on leukocytes from these mouse strains.
  10. Cy is a trademark of GE Healthcare.
  11. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  12. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
555823 Rev. 12
Antibody Details
Down Arrow Up Arrow
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.

555823 Rev. 12
Format Details
Down Arrow Up Arrow
PE-Cy5
PE-Cy5 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496 nm and 566 nm and an acceptor dye, Cy™5, with an emission maximum (Em Max) at 670-nm. PE designed to be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 670-nm (e.g., a 670/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627-640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE-Cy5
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
670 nm
555823 Rev.12
Citations & References
Down Arrow Up Arrow

Development References (5)

  1. 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
  2. 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.
  3. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  4. 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.
  5. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
View All (5) View Less
555823 Rev. 12

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.