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BV605 Mouse Anti-Human CD58
BV605 Mouse Anti-Human CD58
Two-parameter flow cytometric analysis of CD58 expression on human peripheral blood leucocytes. Whole blood was stained with either BD Horizon™ BV605 Mouse IgG2a, κ Isotype Control (Cat. No. 562778; Left Panel) or BD Horizon BV605 Mouse Anti-Human CD58 antibody (Cat. No. 564362; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-parameter contour plots showing the correlated expression of CD58 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals were derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Two-parameter flow cytometric analysis of CD58 expression on human peripheral blood leucocytes. Whole blood was stained with either BD Horizon™ BV605 Mouse IgG2a, κ Isotype Control (Cat. No. 562778; Left Panel) or BD Horizon BV605 Mouse Anti-Human CD58 antibody (Cat. No. 564362; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-parameter contour plots showing the correlated expression of CD58 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals were derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Horizon™
LFA-3; LFA3; Lymphocyte function-associated antigen 3; Ag3
Human (QC Testing), Dog (Tested in Development)
Mouse BALB/c IgG2a, κ
Recombinant Human CD58
Flow cytometry (Routinely Tested)
5 µl
V S029
AB_2738768
Aqueous buffered solution containing ≤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 BD Horizon™ BV605 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV605 were removed.

Recommended Assay Procedures

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).

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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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. 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.
  5. 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 BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
  6. CF™ is a trademark of Biotium, Inc.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. An isotype control should be used at the same concentration as the antibody of interest.
  9. BD Horizon Brilliant Violet 605 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
  10. 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.
564362 Rev. 4
Antibody Details
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1C3

The 1C3 (AICD58.6) monoclonal antibody specifically binds to CD58. CD58 is a a 60-70 kDa glycoprotein member of the immunoglobulin superfamily. CD58, also referred to as the lymphocyte function-associated antigen-3 (LFA-3), has a wide tissue distribution, being expressed on both hematopoietic and non-hematopoietic cells, including endothelial cells and fibroblasts. There are two isoforms of CD58: a glycosylphosphatidylinositol (GPI)-linked form and a transmembrane form. Both isoforms may be expressed on the same cell type. Erythrocytes, however, only express the GPI-linked isoform. CD58 interacts with CD2 during cell adhesion. This binding can enhance antigen-specific T-cell activation. This interaction can also play a role in cell-mediated cytotoxicity.

This antibody is conjugated to BD Horizon BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max of 602-nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20-nm filter set. BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605-nm. Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser.

564362 Rev. 4
Format Details
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BV605
The BD Horizon Brilliant Violet™ 605 (BV605) dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 407-nm and an acceptor dye with an emission maximum (Em Max) at 605-nm. BV605, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 610-nm (e.g., a 610/20-nm bandpass filter). The acceptor dye can be excited by the yellow-green (561-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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BV605
Violet 405 nm
407 nm
605 nm
564362 Rev.4
Citations & References
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Development References (4)

  1. Dengler TJ, Hoffmann JC, Knolle P, et al. Structural and functional epitopes of the human adhesion receptor CD58 (LFA-3). Eur J Immunol. 1992; 22(11):2809-2817. (Clone-specific). View Reference
  2. Framson PE, Cho DH, Lee LY, Hershberg RM. Polarized expression and function of the costimulatory molecule CD58 on human intestinal epithelial cells. Gastroenterology. 1999; 116(5):1054-1062. (Clone-specific: Flow cytometry, Immunofluorescence, Immunoprecipitation, Inhibition). View Reference
  3. 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.
  4. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
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564362 Rev. 4

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.