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 BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; dashed line histogram) or BD Horizon PE-CF594 Mouse Anti-Human CD63 antibody (Cat. No. 565403; solid line histogram). 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.
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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 BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; dashed line histogram) or BD Horizon PE-CF594 Mouse Anti-Human CD63 antibody (Cat. No. 565403; solid line histogram). 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.
Product Details
BD Horizon™
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_2739220
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™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were 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.
- 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.
- When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
- Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
- CF™ is a trademark of Biotium, Inc.
- Texas Red is a registered 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
PE-CF594 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
562292
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565403 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.
This antibody is conjugated to BD Horizon PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg, 610/20-nm filter).
565403 Rev. 1
Format Details
PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
565403 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.
565403 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.
