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BUV395 Mouse Anti-Human CD58
BUV395 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™ BUV395 Mouse IgG2a Isotype Control (Cat. No. 563809; Left Panel) or BD Horizon BUV395 Mouse Anti-Human CD58 antibody (Cat. No. 565460; 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 LSRFortessa™ Cell Analyzer System.
Two-parameter flow cytometric analysis of CD58 expression on human peripheral blood leucocytes. Whole blood was stained with either BD Horizon™ BUV395 Mouse IgG2a Isotype Control (Cat. No. 563809; Left Panel) or BD Horizon BUV395 Mouse Anti-Human CD58 antibody (Cat. No. 565460; 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 LSRFortessa™ Cell Analyzer 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
965
AB_2739248
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 BD Horizon BUV395 under optimum conditions, and unconjugated antibody and free BD Horizon BUV395 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. 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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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. BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
565460 Rev. 2
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.

The antibody was conjugated to BD Horizon BUV395 which has been exclusively developed by BD Biosciences as an optimal dye for use on a 355 nm laser equipped instrument. With an Ex Max at 348 nm  and an Em Max at 395 nm, this dye has virtually no spillover into any other detector. BD Horizon BUV395 can be excited with a 355 nm laser and detected with a 379/28 filter.

565460 Rev. 2
Format Details
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BUV395
The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV395
Ultraviolet 355 nm
348 nm
395 nm
565460 Rev.2
Citations & References
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Development References (5)

  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. (Immunogen: Bioassay, Blocking, ELISA, Flow cytometry, Functional assay, Inhibition, Western blot). 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. Lin G-X, Yang X, Hollemweguer E, et al. Cross-reactivity of CD antibodies in eight animal species. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:519-523.
  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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565460 Rev. 2

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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.