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Alexa Fluor® 647 Hamster Anti-Mouse CD95
Alexa Fluor® 647 Hamster Anti-Mouse CD95

Flow cytometric analysis of CD95 expression on mouse thymocytes. Mouse thymocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either Alexa Fluor® 647 Hamster IgG2, λ 1 isotype control (Cat. No. 562112; dashed line histogram) or Alexa Fluor® 647 Hamster Anti-Mouse CD95 antibody (Cat. No. 563647; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable thymocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of CD95 expression on mouse thymocytes. Mouse thymocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either Alexa Fluor® 647 Hamster IgG2, λ 1 isotype control (Cat. No. 562112; dashed line histogram) or Alexa Fluor® 647 Hamster Anti-Mouse CD95 antibody (Cat. No. 563647; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable thymocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Pharmingen™
Apo-1; Apt1; Fas; FASLG receptor; lpr; TNFR6; Tnfrsf6; TNR6
Mouse (QC Testing)
Armenian Hamster IgG2, λ2
WR19L mouse lymphoma cells transformed with recombinant mouse Fas
Flow cytometry (Routinely Tested)
0.2 mg/ml
14102
AB_2738346
Aqueous buffered solution containing ≤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

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. 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.
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  7. 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.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. 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.
563647 Rev. 1
Antibody Details
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Jo2

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.

563647 Rev. 1
Format Details
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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
563647 Rev.1
Citations & References
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Development References (9)

  1. Enari M, Hug H, Nagata S. Involvement of an ICE-like protease in Fas-mediated apoptosis. Nature. 1995; 375(6526):78-81. (Clone-specific: Functional assay). View Reference
  2. 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, Functional assay). View Reference
  3. 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
  4. Nagata S. Apoptosis regulated by a death factor and its receptor: Fas ligand and Fas. Philos Trans R Soc Lond B Biol Sci. 1994; 345(1313):281-287. (Clone-specific: Functional assay). View Reference
  5. Nagata S. Fas and Fas ligand: a death factor and its receptor. Adv Immunol. 1994; 57:129-144. (Clone-specific: Functional assay). View Reference
  6. Ni R, Tomita Y, Matsuda K, et al. Fas-mediated apoptosis in primary cultured mouse hepatocytes. Exp Cell Res. 1994; 215(2):332-337. (Clone-specific: Functional assay). View Reference
  7. 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
  8. 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
  9. 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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563647 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.