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Flow cytometric analysis of CD95 expression on mouse thymocytes. Thymocytes from a BALB/c mouse were stained with either a BD Horizon™ BV421 Hamster IgG2, λ1 isotype control (Cat. No. 562629; dashed line histogram) or with the BD Horizon™ BV421 Hamster Anti-Mouse CD95 antibody (Cat. No. 562633; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable thymocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.


BD Pharmingen™ BV421 Hamster Anti-Mouse CD95

Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- 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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- Brilliant Violet™ 421 is a trademark of Sirigen.
- Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
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Fas antigen, CD95, is a 45 kDa cell-surface protein which can mediate apoptosis. It belongs to the TNF (tumor necrosis factor)/NGF receptor family. Expression of Fas has been described in the thymus, liver, heart, lung and ovary. Fas plays an important role in the apoptotic process that takes place during development. Monoclonal antibodies recognizing Fas such as Jo2 have cytolytic activity on cells expressing Fas. The cell death stimulated by Fas antibodies is characteristic of apoptosis and suggests that the lethal effects are a result of interaction of antibody with a functional Fas antigen as opposed to complement-mediated lysis.
The Jo2 antibody recognizes mouse Fas. The Jo2 antibody shows cytolytic activity against cell lines expressing mouse Fas by inducing apoptosis. Intraperitoneal injections of Jo2 mAb have been shown to kill mice and induce apoptotic hepatocyte death. Jo2 mAb has been reported to immunoprecipitate mouse Fas as a 45 kDa band from W4 cells. W4 cells are WR19L mouse lymphoma cells transformed with mouse Fas. The difference between the observed MW of Fas and that deduced from its amino acid sequence (Mr 34,971) may be due to glycosylation.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

Development References (5)
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Hiromatsu K, Aoki Y, Makino M, et al. Increased Fas antigen expression in murine retrovirus-induced immunodeficiency syndrome, MAIDS. Eur J Immunol. 1994; 24(10):2446-2451. (Clone-specific: Cytotoxicity, Flow cytometry). View Reference
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Kagi D, Vignaux F, Ledermann B, et al. Fas and perforin pathways as major mechanisms of T cell-mediated cytotoxicity. Science. 1994; 265(5171):528-530. (Clone-specific: Flow cytometry, Functional assay). View Reference
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Ogasawara J, Suda T, Nagata S. Selective apoptosis of CD4+CD8+ thymocytes by the anti-Fas antibody. J Exp Med. 1995; 181(2):485-491. (Clone-specific: Flow cytometry, Functional assay). View Reference
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Ogasawara J, Watanabe-Fukunaga R, Adachi M, et al. Lethal effect of the anti-Fas antibody in mice. Nature. 1993; 364(6440):806-809. (Immunogen: Flow cytometry, Immunoprecipitation). View Reference
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Yang Y, Mercep M, Ware CF, Ashwell JD. Fas and activation-induced Fas ligand mediate apoptosis of T cell hybridomas: inhibition of Fas ligand expression by retinoic acid and glucocorticoids. J Exp Med. 1995; 181(5):1673-1682. (Clone-specific: Flow cytometry). View Reference
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