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BV421 Mouse Anti-Human CD34
BV421 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 BV421 Mouse IgG1 κ Isotype Control (Cat No. 562438, Left Panel) or the BV421 Mouse Anti-Human CD34 antibody (Cat. No. 562577, 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 BV421 Mouse IgG1 κ Isotype Control (Cat No. 562438, Left Panel) or the BV421 Mouse Anti-Human CD34 antibody (Cat. No. 562577, 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 Horizon™
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_2687922
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 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  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. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  7. 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.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
562577 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.

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 BlueTM 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.

562577 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
562577 Rev.3
Citations & References
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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.
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562577 Rev. 3

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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.