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BV421 Mouse Anti-Mouse CD244.2
BV421 Mouse Anti-Mouse CD244.2

Multiparameter flow cytometric analysis using BD OptiBuild™ BV421 Mouse Anti-Mouse CD244.2 antibody (Cat. No. 740015) on live C57BL/6 mouse LAK cells. Flow cytometry was performed using a BD LSRFortessa™ X-20  Flow Cytometer System.

Multiparameter flow cytometric analysis using BD OptiBuild™ BV421 Mouse Anti-Mouse CD244.2 antibody (Cat. No. 740015) on live C57BL/6 mouse LAK cells. Flow cytometry was performed using a BD LSRFortessa™ X-20  Flow Cytometer System.

Product Details
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BD OptiBuild™
CD242.2; 2B4 B6 Alloantigen; Ly90; NAIL; Nmrk; SLAMF4
Mouse (Tested in Development)
Mouse 129, also known as 129/J or 129/SvJ IgG2b, κ
rIL-2-propagated NK1.1+ cells from C57BL/6 mice
Flow cytometry (Qualified)
0.2 mg/ml
18106
AB_2739787
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.
740015 Rev. 2
Antibody Details
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2B4

CD244 is a type I transmembrane glycoprotein that is also known as Natural killer cell receptor 2B4 (NKR2B4), or Signaling lymphocytic activation molecule 4 (SLAMF4). CD244 is a member of the CD2 subset of the immunoglobulin superfamily (CD2 IgSF). It is expressed on all natural killer (NK) cells, IL-2-activated NK (LAK) cells, committed progenitors of NK cells, and a subset of T lymphocytes which mediate non-MHC-restricted cytoxicity, including dendritic epidermal T cells. The 2B4 monoclonal antibody specifically binds to CD244.2, the 2B4 alloantigen which is expressed on C57BL/6 and C58/J mice, but not in most strains tested (A/J, AKR, BALB/c, CBA/J, CBA/N, C3H/He, C57BR, DBA/1, DBA/2, NZB, SJL/J, 129). At least two isoforms of CD244.2 protein, products of alternative splicing of hnRNA, are expressed on IL-2-activated C57BL/6 NK cells. They differ only in their cytoplasmic domains; with 2B4L (150-aa cytoplasmic tail) having inhibitory activity and 2B4S (93-aa tail) being stimulatory. The extracellular domain of CD244 is a ligand for another CD2 IgSF member, CD48. 2B4 antibody activates lytic and secretory functions of IL-2-cultured NK cells and CD244.2+ T cells.

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.

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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
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Citations & References
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Development References (13)

  1. Brown MH, Boles K, van der Merwe PA, Kumar V, Mathew PA, Barclay AN. 2B4, the natural killer and T cell immunoglobulin superfamily surface protein, is a ligand for CD48. J Exp Med. 1998; 188(11):2083-2090. (Biology). View Reference
  2. Davis SJ, van der Merwe PA. The structure and ligand interactions of CD2: implications for T-cell function. Immunol Today. 1996; 17(4):177-187. (Biology). View Reference
  3. Garni-Wagner BA, Purohit A, Mathew PA, Bennett M, Kumar V. A novel function-associated molecule related to non-MHC-restricted cytotoxicity mediated by activated natural killer cells and T cells. J Immunol. 1993; 151(1):60-70. (Biology). View Reference
  4. Kubota K, Katoh H, Muguruma K, Koyama K. Characterization of a surface membrane molecule expressed by natural killer cells in most inbred mouse strains: monoclonal antibody C9.1 identifies an allelic form of the 2B4 antigen. Immunology. 1999; 96(3):491-497. (Biology). View Reference
  5. Kumaresan PR, Stepp SE, Bennett M, Kumar V, Mathew PA. Molecular cloning of transmembrane and soluble forms of a novel rat natural killer cell receptor related to 2B4. Immunogenetics. 2000; 51(4-5):306-313. (Biology). View Reference
  6. Latchman Y, McKay PF, Reiser H. Identification of the 2B4 molecule as a counter-receptor for CD48. J Immunol. 1998; 161(11):5809-5812. (Biology). View Reference
  7. Mathew PA, Garni-Wagner BA, Land K, et al. Cloning and characterization of the 2B4 gene encoding a molecule associated with non-MHC-restricted killing mediated by activated natural killer cells and T cells. J Immunol. 1993; 151(10):5328-5337. (Biology). View Reference
  8. Rosmaraki EE, Douagi I, Roth C, Colucci F, Cumano A, Di Santo JP. Identification of committed NK cell progenitors in adult murine bone marrow. Eur J Immunol. 2001; 31(6):1900-1909. (Biology). View Reference
  9. Schatzle JD, Sheu S, Stepp SE, Mathew PA, Bennett M, Kumar V. Characterization of inhibitory and stimulatory forms of the murine natural killer cell receptor 2B4. Proc Natl Acad Sci U S A. 1999; 96(7):3870-3875. (Biology). View Reference
  10. Schuhmachers G, Ariizumi K, Mathew PA, Bennett M, Kumar V, Takashima A. 2B4, a new member of the immunoglobulin gene superfamily, is expressed on murine dendritic epidermal T cells and plays a functional role in their killing of skin tumors. J Invest Dermatol. 1995; 105(4):592-596. (Biology). View Reference
  11. Schuhmachers G, Ariizumi K, Mathew PA, Bennett M, Kumar V, Takashima A. Activation of murine epidermal gamma delta T cells through surface 2B4. Eur J Immunol. 1995; 25(4):1117-1120. (Biology). View Reference
  12. Sentman CL, Hackett J Jr, Moore TA, Tutt MM, Bennett M, Kumar V. Pan natural killer cell monoclonal antibodies and their relationship to the NK1.1 antigen. Hybridoma. 1989; 8(6):605-614. (Immunogen). View Reference
  13. Stepp SE, Schatzle JD, Bennett M, Kumar V, Mathew PA. Gene structure of the murine NK cell receptor 2B4: presence of two alternatively spliced isoforms with distinct cytoplasmic domains. Eur J Immunol. 1999; 29(8):2392-2399. (Biology). View Reference
View All (13) View Less
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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.