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PE-Cy™7 Mouse Anti-Human CD34
PE-Cy™7 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™7 Mouse IgG1 κ Isotype Control (Cat No. 557872, Left Panel) or the PE-Cy™7-Mouse Anti-Human CD34 antibody (Cat. No. 560710, 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™7 Mouse IgG1 κ Isotype Control (Cat No. 557872, Left Panel) or the PE-Cy™7-Mouse Anti-Human CD34 antibody (Cat. No. 560710, 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
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BD Pharmingen™
gp105-120; My10; Hematopoietic progenitor cell antigen CD34
Human (QC Testing)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl
V MA27, VI E004
947
AB_1727470
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-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 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. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
  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-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Cy is a trademark of GE Healthcare.
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560710 Rev. 3
Antibody Details
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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.

560710 Rev. 3
Format Details
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PE-Cy7
PE-Cy7 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™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-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.
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PE-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
560710 Rev.3
Citations & References
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Development References (6)

  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. 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.
  5. 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.
  6. 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.
View All (6) View Less
560710 Rev. 3

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