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 wash, the fixed platelets were stained with either Alexa Fluor® 647 Mouse Anti-Human CD63 antibody (Cat. No. 561983; solid line histogram) or with an Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; dashed line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of platelets. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
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 wash, the fixed platelets were stained with either Alexa Fluor® 647 Mouse Anti-Human CD63 antibody (Cat. No. 561983; solid line histogram) or with an Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; dashed line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of platelets. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
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)
5 µl
V P036
967
AB_10897006
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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- 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).
- 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.
- 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
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557714
561983 Rev. 2
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.
561983 Rev. 2
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
561983 Rev.2
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. (Biology). 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. (Biology). 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.
561983 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.
