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PE Mouse Anti-Mouse TCR Vδ6B

BD Pharmingen™ PE Mouse Anti-Mouse TCR Vδ6B

Clone F4.22.rMAb (also known as Anti-Vδ6λ12) (RUO)

PE Mouse Anti-Mouse TCR Vδ6B

Flow cytometric analysis of TCR Vδ6B expression on Mouse intraepithelial lymphocytes (IELs). C57BL/6 Mouse IELs were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141). The cells were then stained with BD OptiBuild™ R718 Hamster Anti-Mouse γδ T-Cell Receptor (Cat No. 751919), FITC Hamster Anti-Mouse CD3e (Cat. No. 553062) and with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Mouse TCR Vδ6B antibody (Cat. No. 570738/570739; Right Plot) at 0.5 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of TCR Vδ6B (or Ig Isotype control staining) versus TCRγδ was derived from gated events with the forward and side light- scatter characteristics of viable (DAPI-negative) CD3-positive IELs. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.

Flow cytometric analysis of TCR Vδ6B expression on Mouse intraepithelial lymphocytes (IELs). C57BL/6 Mouse IELs were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141). The cells were then stained with BD OptiBuild™ R718 Hamster Anti-Mouse γδ T-Cell Receptor (Cat No. 751919), FITC Hamster Anti-Mouse CD3e (Cat. No. 553062) and with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-Mouse TCR Vδ6B antibody (Cat. No. 570738/570739; Right Plot) at 0.5 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of TCR Vδ6B (or Ig Isotype control staining) versus TCRγδ was derived from gated events with the forward and side light- scatter characteristics of viable (DAPI-negative) CD3-positive IELs. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Pharmingen™
TCR Variable delta 6B; TCR Vd6b; TCR Vδ6λ12
Mouse (QC Testing)
Mouse IgG1, κ
Mouse γδ T Cell Hybridoma Cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
110371
AB_3686012
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. For U.S. patents that may apply, see bd.com/patents.
570739 Rev. 1
Antibody Details
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F4.22.rMAb

F4.22.RMAB is a recombinant form of the F4.22 monoclonal antibody. The F4.22 antibody specifically recognizes TCR Vδ6B subfamily members that have more than 90% identity at the nucleotide level with the previously described pλ12 and is also referred to as Anti-Vδ6λ12. The F4.22 antibody recognizes two members of the Vδ6/ADV subfamily expressed in C57BL/6 (B6) mice and at least one subfamily member expressed in DBA/2 mice. The F4.22 antibody does not recognize any TCR γδ T cell hybridomas expressing different TCR Vδ chains including other members of the TCR Vδ6 subfamily. The F4.22 antibody stains sizable proportions of Vγ1+, Vγ4+, and Vγ7+ γδ T cells prepared from the thymus, spleen, lymph nodes and intraepithelial lymphocytes (IELs) and T cell hybridomas derived from B6 mice. It likewise stains Vγ1+ and Vγ7+ γδ T cells from C57BL/10 and B10.D2 that share the same TCRδ haplotype. It does not stain any γδ T cells from BALB/c, C3H/HeJ, CBA/J, 129/Sv and FVB/N mice.

570739 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 561 nm
496 nm, 566 nm
576 nm
570739 Rev.1
Citations & References
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View product citations for antibody "570739" on CiteAb

Development References (7)

  1. Azuara V, Grigoriadou K, Lembezat MP, Nagler-Anderson C, Pereira P. Strain-specific TCR repertoire selection of IL-4-producing Thy-1 dull gamma delta thymocytes. Eur J Immunol. 2001; 31(1):205-214. (Biology). View Reference
  2. Boucontet L, Grana M, Alzari PM, Pereira P. Mechanisms determining cell membrane expression of different gammadelta TCR chain pairings.. Eur J Immunol. 2009; 39(7):1937-46. (Biology). View Reference
  3. Chien YH, Iwashima M, Kaplan KB, Elliott JF, Davis MM. A new T-cell receptor gene located within the alpha locus and expressed early in T-cell differentiation.. Nature. 327(6124):677-82. (Biology). View Reference
  4. French JD, Roark CL, Born WK, O'Brien RL. Gammadelta T lymphocyte homeostasis is negatively regulated by beta2-microglobulin. J Immunol. 2009; 182(4):1892-1900. (Clone-specific: Flow cytometry). View Reference
  5. Pereira P, Berthault C, Burlen-Defranoux O, Boucontet L. Critical role of TCR specificity in the development of Vγ1Vδ6.3+ innate NKTγδ cells. J Immunol. 2013; 191(4):1716-1723. (Clone-specific: Flow cytometry). View Reference
  6. Pereira P, Hermitte V, Lembezat MP, Boucontet L, Azuara V, Grigoriadou K. Developmentally regulated and lineage-specific rearrangement of T cell receptor Valpha/delta gene segments.. Eur J Immunol. 2000; 30(7):1988-97. (Immunogen: Flow cytometry). View Reference
  7. Zeng W, O'Brien RL, Born WK, Huang Y. Characterization of Mouse γδ T Cell Subsets in the Setting of Type-2 Immunity.. Methods Mol Biol. 2018; 1799:135-151. (Clone-specific: Flow cytometry). View Reference
View All (7) View Less
570739 Rev. 1

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