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BV421 Hamster Anti-Mouse γδ T-Cell Receptor
BV421 Hamster Anti-Mouse γδ T-Cell Receptor

Multicolor flow cytometric analysis of γδ TCR expression on mouse peripheral T lymphocytes. C57BL/6 lymph node cells were incubated simultaneously with PE Rat anti-Mouse CD3 Molecular Complex (Cat. No. 555275/561799) and with either BV421 Hamster IgG2, κ Isotype Control (Cat. No. 562612, left panel) or BV421 Hamster Anti-Mouse γδ T-Cell Receptor (Cat. No. 562892, right panel). Two-color flow cytometric dot plots showing the correlated expression of γδ T-Cell Receptor (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and light-scattering characteristics of viable lymphocytes. Flow cytometry was performed on a BD™ LSR II.

Multicolor flow cytometric analysis of γδ TCR expression on mouse peripheral T lymphocytes. C57BL/6 lymph node cells were incubated simultaneously with PE Rat anti-Mouse CD3 Molecular Complex (Cat. No. 555275/561799) and with either BV421 Hamster IgG2, κ Isotype Control (Cat. No. 562612, left panel) or BV421 Hamster Anti-Mouse γδ T-Cell Receptor (Cat. No. 562892, right panel). Two-color flow cytometric dot plots showing the correlated expression of γδ T-Cell Receptor (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and light-scattering characteristics of viable lymphocytes. Flow cytometry was performed on a BD™ LSR II.

Product Details
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BD Horizon™
Tcrd; T-cell receptor delta chain; Tcr delta
Mouse (QC Testing)
Armenian Hamster IgG2, κ
C57BL/6 Mouse Intestinal Intraepithelial Lymphocytes
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2737871
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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

For flow cytometry of cell suspensions from peripheral lymphoid tissues, it is recommended that multicolor staining be performed to distinguish Tlymphocytes from non-T cells.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes 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 Brilliant Stain Buffer (Cat. No. 563794/566349) or the Brilliant Stain Buffer Plus (Cat. No. 566385).

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. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. 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.
  6. 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.
  7. 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.
  8. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  10. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
562892 Rev. 2
Antibody Details
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GL3

The GL3 monoclonal antibody specifically binds to a common epitope of the δ chain of the T-cell Receptor (TCR) complex on γδ TCR-expressing T lymphocytes and NK-T cells of all mouse strains tested. It does not react with αβ TCR-bearing T cells. In the mouse, cells expressing the γδ TCR are found in the thymus, intestinal epithelium, epidermis, dermis, pulmonsry epithelium, peritoneum, liver, and peripheral lymphoid organs.

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.

562892 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
562892 Rev.2
Citations & References
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Development References (15)

  1. Goodman T, LeCorre R, Lefrancois L. A T-cell receptor gamma delta-specific monoclonal antibody detects a V gamma 5 region polymorphism. Immunogenetics. 1992; 35(1):65-68. (Clone-specific: Flow cytometry). View Reference
  2. Goodman T, Lefrancois L. Intraepithelial lymphocytes. Anatomical site, not T cell receptor form, dictates phenotype and function. J Exp Med. 1989; 170(5):1569-1581. (Clone-specific: Flow cytometry, Immunoprecipitation). View Reference
  3. Kaufmann SH, Blum C, Yamamoto S. Crosstalk between alpha/beta T cells and gamma/delta T cells in vivo: activation of alpha/beta T-cell responses after gamma/delta T-cell modulation with the monoclonal antibody GL3. Proc Natl Acad Sci U S A. 1993; 90(20):9620-9624. (Clone-specific: Flow cytometry, In vivo exacerbation). View Reference
  4. King DP, Hyde DM, Jackson KA, et al. Cutting edge: protective response to pulmonary injury requires gamma delta T lymphocytes. J Immunol. 1999; 162(9):5033-5036. (Biology: Flow cytometry). View Reference
  5. Lefrancois L, Barrett TA, Havran WL, Puddington L. Developmental expression of the alpha IEL beta 7 integrin on T cell receptor gamma delta and T cell receptor alpha beta T cells. Eur J Immunol. 1994; 24(3):635-640. (Clone-specific: Immunohistochemistry). View Reference
  6. Lefrancois L. Phenotypic complexity of intraepithelial lymphocytes of the small intestine. J Immunol. 1991; 147(6):1746-1751. (Clone-specific: Flow cytometry). View Reference
  7. MacDonald HR, Schreyer M, Howe RC, Bron C. Selective expression of CD8 alpha (Ly-2) subunit on activated thymic gamma/delta cells. Eur J Immunol. 1990; 20(4):927-930. (Biology: Flow cytometry). View Reference
  8. Nakazawa S, Brown AE, Maeno Y, Smith CD, Aikawa M. Malaria-induced increase of splenic gamma delta T cells in humans, monkeys, and mice. 1994; 79(3):391-398. (Clone-specific: Immunohistochemistry). View Reference
  9. Shinohara K, Ikarashi Y, Maruoka H, et al. Functional and phenotypical characteristics of hepatic NK-like T cells in NK1.1-positive and -negative mouse strains. Eur J Immunol. 1999; 29(6):1871-1878. (Biology: Flow cytometry). View Reference
  10. Skeen MJ, Ziegler HK. Induction of murine peritoneal gamma/delta T cells and their role in resistance to bacterial infection. J Exp Med. 1993; 178(3):971-984. (Biology: Flow cytometry). View Reference
  11. Tamaki K, Yasaka N, Chang CH, et al. Identification and characterization of novel dermal Thy-1 antigen-bearing dendritic cells in murine skin. J Invest Dermatol. 1996; 106(3):571-575. (Biology: Flow cytometry). View Reference
  12. Tigelaar RE, Lewis JM, Bergstresser PR. TCR gamma/delta+ dendritic epidermal T cells as constituents of skin-associated lymphoid tissue. J Invest Dermatol. 1990; 94(6):58S-63S. (Biology). View Reference
  13. Vicari AP, Mocci S, Openshaw P, O'Garra A, Zlotnik A. Mouse gamma delta TCR+NK1.1+ thymocytes specifically produce interleukin-4, are major histocompatibility complex class I independent, and are developmentally related to alpha beta TCR+NK1.1+ thymocytes. Eur J Immunol. 1996; 26(7):1424-1429. (Biology: Flow cytometry). View Reference
  14. Yanez DM, Batchelder J, van der Heyde HC, Manning DD, Weidanz WP. Gamma delta T-cell function in pathogenesis of cerebral malaria in mice infected with Plasmodium berghei ANKA. Infect Immun. 1999; 67(1):446-448. (Clone-specific: Depletion). View Reference
  15. van der Heyde HC, Elloso MM, Chang WL, Kaplan M, Manning DD, Weidanz WP. Gamma delta T cells function in cell-mediated immunity to acute blood-stage Plasmodium chabaudi adami malaria. J Immunol. 1995; 154(8):3985-3990. (Clone-specific: Depletion). View Reference
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562892 Rev. 2

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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.