Multiparameter flow cytometric analysis of CD81 expression on human peripheral blood leucocytes. Human whole blood was stained with either PerCP-Cy™5.5 Mouse IgG1, κ Isotype Control (Cat. No. 550795; Left Panel) or PerCP-Cy™5.5 Mouse Anti-Human CD81 antibody (Cat. No. 565430; Right Panel). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric contour plots showing the correlated expression of CD81 (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.
Multiparameter flow cytometric analysis of CD81 expression on human peripheral blood leucocytes. Human whole blood was stained with either PerCP-Cy™5.5 Mouse IgG1, κ Isotype Control (Cat. No. 550795; Left Panel) or PerCP-Cy™5.5 Mouse Anti-Human CD81 antibody (Cat. No. 565430; Right Panel). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric contour plots showing the correlated expression of CD81 (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 Pharmingen™
TAPA1; Tetraspanin-28; Tspan-28; TSPAN28; CVID6; S5.7; M38
Human (QC Testing), Rhesus, Cynomolgus, Baboon, Dog, Rabbit, Cat, Horse (Tested in Development)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl
V B070
975
AB_2739235
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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
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.
- 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.
- PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
- PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- 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.
- Cy is a trademark of GE Healthcare.
- 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 CE_IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
PerCP-Cy™5.5 Mouse IgG1 κ Isotype Control RUO
Size
0.1 mg
Cat No.
550795
565430 Rev. 3
Antibody Details
JS-81
The JS-81 monoclonal antibody specifically binds to CD81, which is also known as, Target of the antiproliferative antibody 1 (TAPA1, TAPA-1), or Tetraspanin-28 (Tspan-28/TSPAN28). CD81 is an ~26 kDa transmembrane protein that belongs to the tetraspanin (TM4SF) family. It is involved in cell growth and signal transduction. CD81 has a very broad cellular distribution, being expressed on cells of hematopoietic, neuroectodermal and mesenchymal origin. In hematopoietic cells, the CD81 antigen is expressed on B and T lymphocytes, NK cells, thymocytes, eosinophils, germinal center follicular dendritic cells, and to a variable extent on monocytes. The CD81 antigen is not expressed on neutrophils, platelets, or erythrocytes. CD81-specific antibodies have been shown to have anti-proliferative effects on different lymphoid cell lines, particularly those derived from large cell lymphomas. They are also reported to induce homotypic cell aggregation. Immunoprecipitation studies reveal that CD81 is a component of a multimolecular complex of CD19, CD21, and CD225 that is involved in the activation and control of B cell growth.
565430 Rev. 3
Format Details
PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-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.
PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
565430 Rev.3
Citations & References
Development References (8)
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Bradbury LE, Kansas GS, Levy S, Evans RL, Tedder TF. The CD19/CD21 signal transducing complex of human B lymphocytes includes the target of antiproliferative antibody-1 and Leu-13 molecules. J Immunol. 1992; 149(9):2841-2850. (Biology). View Reference
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Carloni V, Mazzocca A, Ravichandran KS. Tetraspanin CD81 is linked to ERK/MAPKinase signaling by Shc in liver tumor cells. Oncogene. 2004; 23(8):1566-1574. (Clone-specific: Flow cytometry, Immunofluorescence, Immunoprecipitation). View Reference
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Levy S, Todd SC, Maecker HT. CD81 (TAPA-1): a molecule involved in signal transduction and cell adhesion in the immune system. Annu Rev Immunol. 1998; 16:89-109. (Clone-specific). View Reference
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Lin G-X, Yang X, Hollemweguer E, et al. Cross-reactivity of CD antibodies in eight animal species. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:519-523.
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Mazzocca A, Sciammetta SC, Carloni V, et al. Binding of hepatitis C virus envelope protein E2 to CD81 up-regulates matrix metalloproteinase-2 in human hepatic stellate cells. J Biol Chem. 2005; 280(12):11329-11339. (Clone-specific: Blocking, Flow cytometry, Functional assay, Inhibition, Neutralization). View Reference
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Oren R, Takahashi S, Doss C, Levy R, Levy S. TAPA-1, the target of an antiproliferative antibody, defines a new family of transmembrane proteins. Mol Cell Biol. 1990; 10(8):4007-4015. (Biology). View Reference
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Schick MR, Levy S. The TAPA-1 molecule is associated on the surface of B cells with HLA-DR molecules. J Immunol. 1993; 151(8):4090-4097. (Biology). View Reference
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Tedder TF, Wagner N, Engel P. CD81 Workshop report. In: 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; 1995:684-688.
565430 Rev. 3
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.
