Flow cytometric analysis of CD58 expression on human peripheral blood lymphocytes. Whole blood was stained with either Alexa Fluor® 647 Mouse Anti-Human CD58 antibody (Cat. No. 563567; solid line histogram) or Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 557715; dashed line histogram). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD58 expression on human peripheral blood lymphocytes. Whole blood was stained with either Alexa Fluor® 647 Mouse Anti-Human CD58 antibody (Cat. No. 563567; solid line histogram) or Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 557715; dashed line histogram). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
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_2738280
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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
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).
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control RUO
Size
100 Tests
Cat No.
557715
563567 Rev. 1
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.
563567 Rev. 1
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
563567 Rev.1
Citations & References
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. (Clone-specific). 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.
563567 Rev. 1
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.
