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BV421 Hamster Anti-Mouse CD178
BV421 Hamster Anti-Mouse CD178

Multiparameter flow cytometric analysis using BD OptiBuild™ BV421 Hamster Anti-Mouse CD178 antibody (Cat. No. 740054) on live enriched T cells from BALB/c mouse spleen stimulated by purified plate bound CD3e antibody. Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.

Multiparameter flow cytometric analysis using BD OptiBuild™ BV421 Hamster Anti-Mouse CD178 antibody (Cat. No. 740054) on live enriched T cells from BALB/c mouse spleen stimulated by purified plate bound CD3e antibody. Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.

Product Details
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BD OptiBuild™
Fas Ligand, FasL; Fas-Lg; CD95 Ligand; CD95L; Tnfsf6; APT1LG1; gld
Mouse (Tested in Development)
Armenian Hamster IgG1, κ
Mouse FasL-transfected cells
Flow cytometry (Qualified)
0.2 mg/ml
AB_2870633
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 with BD Horizon BV421 under optimal conditions that minimize unconjugated dye and antibody.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

Product Notices

  1. This antibody was developed for use in flow cytometry.
  2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  3. Researchers should determine the optimal concentration of this reagent for their individual applications.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  10. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
740054 Rev. 2
Antibody Details
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MFL3

The MFL3 monoclonal antibody specifically binds to CD178 (Fas Ligand, CD95 Ligand) on all strains tested. In the mouse, Fas Ligand is expressed on activated T cell lines and in spleen, testis, and eye. FasL mRNA has been demonstrated at various levels in bone marrow, thymus, spleen, lymph node, lung, small intestine, testis, and uterus. Moreover, T-cell activators, but not B-cell activators, enhanced the expression of FasL mRNA in splenocytes; and FasL mRNA was restricted to the T-cell lineage among a panel of cell lines from lymphoid tissues. Fas Ligand is not functional in mice homozygous for the gld (generalized lympho-proliferative disease) mutation; these mice cannot limit the expansion of activated lymphocytes and develop autoimmune disease. Fas Ligand is a member of the TNF/NGF family, which binds to CD95 (Fas), inducing apoptotic cell death. This Fas/Fas Ligand interaction is believed to participate in T-cell development, the regulation of immune responses, and cell-mediated cytotoxic mechanisms. There is mounting evidence that Fas Ligand is also proinflammatory, mediating neutrophil extravasation and chemotaxis. Fas Ligand is released from the surface of transfectant cells by metalloproteinases, and the soluble Fas Ligand may block the activities of the membrane-bound molecule. The MFL3 mAb has been  reported to efficiently inhibit the cytotoxicity of mouse Fas Ligand-transfected cells against human Fas-transfected cells. This hamster mAb to a mouse leukocyte antigen does not cross-react with rat leukocytes.

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.

740054 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
740054 Rev.2
Citations & References
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Development References (17)

  1. Bellgrau D, Gold D, Selawry H, Moore J, Franzusoff A, Duke RC. A role for CD95 ligand in preventing graft rejection. Nature. 1995; 377(6550):630-632. (Biology). View Reference
  2. Brunner T, Mogil RJ, LaFace D, et al. Cell-autonomous Fas (CD95)/Fas-ligand interaction mediates activation-induced apoptosis in T-cell hybridomas. Nature. 1995; 373:441-444. (Biology). View Reference
  3. Fuller CL, Ravichandran KS, Braciale VL. Phosphatidylinositol 3-kinase-dependent and -independent cytolytic effector functions. J Immunol. 1999; 162(11):6337-6340. (Clone-specific: Inhibition). View Reference
  4. Griffith TS, Brunner T, Fletcher SM, Green DR, Ferguson TA. Fas ligand-induced apoptosis as a mechanism of immune privilege. Science. 1995; 270(5239):1189-1192. (Biology). View Reference
  5. Griffith TS, Ferguson TA. The role of FasL-induced apoptosis in immune privilege. Immunol Today. 1997; 18(5):240-244. (Biology). View Reference
  6. Hohlbaum AM, Moe S, Marshak-Rothstein A. Opposing effects of transmembrane and soluble Fas ligand expression on inflammation and tumor cell survival. J Exp Med. 2000; 191(7):1209-1220. (Biology). View Reference
  7. Ju ST, Panka DJ, Cui H, et al. Fas(CD95)/FasL interactions required for programmed cell death after T-cell activation. Nature. 1995; 373(6513):444-448. (Biology). View Reference
  8. Kayagaki N, Yamaguchi N, Nagao F, et al. Polymorphism of murine Fas ligand that affects the biological activity.. Proc Natl Acad Sci USA. 1997; 94(8):3914-9. (Immunogen: Inhibition). View Reference
  9. Kojima H, Shinohara N, Hanaoka S, et al. Two distinct pathways of specific killing revealed by perforin mutant cytotoxic T lymphocytes. Immunity. 1994; 1(5):357-364. (Biology). View Reference
  10. Lau HT, Yu M, Fontana A, Stoeckert CJ Jr. Prevention of islet allograft rejection with engineered myoblasts expressing FasL in mice. Science. 1996; 273(5271):109-112. (Biology). View Reference
  11. Lynch DH, Ramsdell F, Alderson MR. Fas and FasL in the homeostatic regulation of immune responses. Immunol Today. 1995; 16(12):569-574. (Biology). View Reference
  12. Ramsdell F, Seaman MS, Miller RE, Picha KS, Kennedy MK, Lynch DH. Differential ability of Th1 and Th2 T cells to express Fas ligand and to undergo activation-induced cell death. Int Immunol. 1994; 6(10):1545-1553. (Biology). View Reference
  13. Schneider P, Holler N, Bodmer JL, et al. Conversion of membrane-bound Fas(CD95) ligand to its soluble form is associated with downregulation of its proapoptotic activity and loss of liver toxicityq. J Exp Med. 1998; 187(8):1205-1213. (Biology). View Reference
  14. Smith CA, Farrah T, Goodwin RG. The TNF receptor superfamily of cellular and viral proteins: activation, costimulation, and death. Cell. 1994; 76(6):959-962. (Biology). View Reference
  15. Suda T, Okazaki T, Naito Y, et al. Expression of the Fas ligand in cells of T cell lineage. J Immunol. 1995; 154(8):3806-3813. (Biology). View Reference
  16. Takahashi T, Tanaka M, Brannan CI, et al. Generalized lymphoproliferative disease in mice, caused by a point mutation in the Fas ligand. Cell. 1994; 76(6):969-976. (Biology). View Reference
  17. Vignaux F, Vivier E, Malissen B, Depraetere V, Nagata S, Golstein P. TCR/CD3 coupling to Fas-based cytotoxicity. J Exp Med. 1995; 181(2):781-786. (Biology). View Reference
View All (17) View Less
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.