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BD Horizon™ BV421 Mouse Anti-Human CD58
Clone 1C3 (also known as AICD58.6) (RUO)
Two-parameter flow cytometric analysis of CD58 expression on human peripheral blood leucocytes. Whole blood was stained with either BD Horizon™ BV421 Mouse IgG2a Isotype Control (Cat. No. 562439; Left Panel) or BD Horizon BV421 Mouse Anti-Human CD58 antibody (Cat. No. 564363; 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™ BV421 Mouse IgG2a Isotype Control (Cat. No. 562439; Left Panel) or BD Horizon BV421 Mouse Anti-Human CD58 antibody (Cat. No. 564363; 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™ BV421 Mouse IgG2a Isotype Control (Cat. No. 562439; Left Panel) or BD Horizon BV421 Mouse Anti-Human CD58 antibody (Cat. No. 564363; 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™ BV421 Mouse IgG2a Isotype Control (Cat. No. 562439; Left Panel) or BD Horizon BV421 Mouse Anti-Human CD58 antibody (Cat. No. 564363; 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
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_2738769
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 BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 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
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- 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.
- 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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Lysing Solution 10X Concentrate IVD
Size
100 ML
Cat No.
349202
BV421 Mouse IgG2a, k Isotype Control RUO
Size
50 µg
Cat No.
562439
564363 Rev. 2
Antibody Details
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™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
564363 Rev. 2
Format Details
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.
BV421
407 nm
423 nm
564363 Rev.2
Citations & References
View product citations for antibody "564363" on CiteAb
Development References (4)
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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
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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
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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.
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
564363 Rev. 2
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.