Flow cytometric analysis of CD63 expression on human peripheral blood platelets. Platelets were isolated from fresh whole blood and activated by Thrombin (Sigma-Aldrich, Cat. No. T8885), and then fixed with 2% formaldehyde. After washing, the fixed platelets were stained with either Biotin Mouse IgG1 κ Isotype Control (Cat. No. 555747; dashed line histogram) or Biotin Mouse Anti-Human CD63 antibody (Cat. No. 567729; solid line histogram) at 0.5 µg/test, followed by PE Streptavidin (Cat. No. 554061). The fluorescence histogram showing CD63 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact platelets. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System. Data shown on this Technical Data Sheet are not lot-specific.
Flow cytometric analysis of CD63 expression on human peripheral blood platelets. Platelets were isolated from fresh whole blood and activated by Thrombin (Sigma-Aldrich, Cat. No. T8885), and then fixed with 2% formaldehyde. After washing, the fixed platelets were stained with either Biotin Mouse IgG1 κ Isotype Control (Cat. No. 555747; dashed line histogram) or Biotin Mouse Anti-Human CD63 antibody (Cat. No. 567729; solid line histogram) at 0.5 µg/test, followed by PE Streptavidin (Cat. No. 554061). The fluorescence histogram showing CD63 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact platelets. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System. Data shown on this Technical Data Sheet are not lot-specific.
Product Details
BD Pharmingen™
LAMP-3; ME491; MLA-1; Granulophysin; Ptgr40; NGA; gp55
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse BALB/c IgG1, κ
Human Splenic Adherent Cells
Flow cytometry (Routinely Tested)
0.5 mg/ml
V P036
967
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed. Store undiluted at 4°C.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
567729 Rev. 1
Antibody Details
H5C6
The H5C6 monoclonal antibody specifically binds to CD63. CD63 is a 53 kDa, type III lysosomal glycoprotein, expressed on activated platelets, monocytes and macrophages. This molecule is also referred to in the literature as LIMP, gp55, melanoma-associated antigen ME491, Pltgp40, LAMP-3 and is a member of the tetraspan transmembrane 4 superfamily (TM4SF). It is widely expressed on surface and in the cytoplasm of various hematopoietic (monocytes, macrophages) and non-hematopoietic (endothelium, fibroblasts, osteoclasts, smooth muscle) cells. CD63 plays roles in mediating cellular adhesion and motility.
567729 Rev. 1
Format Details
Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
567729 Rev.1
Citations & References
Development References (4)
-
Azorsa DO, Hyman JA, Hildreth JE. CD63/Pltgp40: a platelet activation antigen identical to the stage-specific, melanoma-associated antigen ME491. Blood. 1991; 78(2):280-284. (Clone-specific: Immunoprecipitation). View Reference
-
Hildreth JE, Derr D, Azorsa DO. Characterization of a novel self-associating Mr 40,000 platelet glycoprotein. Blood. 1991; 77(1):121-132. (Immunogen: Flow cytometry, Immunohistochemistry, Immunoprecipitation, Radioimmunoassay). View Reference
-
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.
-
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.
567729 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.
