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PE Rat Anti-Mouse Vα 8.3 TCR
PE Rat Anti-Mouse Vα 8.3 TCR

Two-color analysis of the expression of Vα 8.3 TCR on peripheral lymphocytes. C57BL/6 lymph node cells were incubated simultaneously with PE-conjugated B21.14, FITC-conjugated anti-mouse CD4 RM4-5 (Cat. No. 553046/553047), and FITC-conjugated anti-mouse CD8a 53-6.7 (Cat. No. 553030/553031) monoclonal antibodies. Flow cytometry was performed on a BD FACScan™ flow cytometry system.

Two-color analysis of the expression of Vα 8.3 TCR on peripheral lymphocytes. C57BL/6 lymph node cells were incubated simultaneously with PE-conjugated B21.14, FITC-conjugated anti-mouse CD4 RM4-5 (Cat. No. 553046/553047), and FITC-conjugated anti-mouse CD8a 53-6.7 (Cat. No. 553030/553031) monoclonal antibodies. Flow cytometry was performed on a BD FACScan™ flow cytometry system.

Product Details
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BD Pharmingen™
Mouse (QC Testing)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG1, κ
Soluble αβ TCR from mouse cytotoxic T-cell clone CW3/1.1
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_394821
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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

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

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. 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.
553377 Rev. 10
Antibody Details
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B21.14

The B21.14 monoclonal antibody specifically recognizes some members of the Vα 8 T-cell Receptor (TCR) subfamily of mice having the a, b, c, and d haplotypes of the Tcra gene complex (e.g., all strains tested). It recognizes an epitope in the CDR1 of Vα 8.3, but not Vα 8.2, TCR subfamily member, as does the KT50 mAb (Cat. No. 557096, for the biotinylated antibody). Site-directed mutagenesis has identified three amino acids which are necessary for antibody reactivity and which are unique to Vα 8.3 among the five functional Vα 8 TCR subfamily members. On a common H-2K[k] background and using the KT50 mAb, the frequency of Vα 8.3 TCR-bearing T lymphocytes has been demonstrated to be higher in Tcra[a/a] mice than in Tcra[a/b] mice. Furthermore, studies of congenic strains suggest that CD8+ Vα 8.3 TCR-bearing T lymphocytes undergo negative selection in mice expressing MHC class I antigens of the H-2[d] haplotype.

553377 Rev. 10
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 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
553377 Rev.10
Citations & References
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Development References (4)

  1. Brodnicki TC, Holman PO, Kranz DM. Reactivity and epitope mapping of single-chain T cell receptors with monoclonal antibodies. Mol Immunol. 1996; 33(3):253-263. (Biology). View Reference
  2. Necker A, Rebai N, Matthes M, et al. Monoclonal antibodies raised against engineered soluble mouse T cell receptors and specific for V alpha 8-, V beta 2- or V beta 10-bearing T cells. Eur J Immunol. 1991; 21(12):3035-3040. (Immunogen). View Reference
  3. Tomonari K, Fairchild S, Rosenwasser OA. Influence of viral superantigens on V beta- and V alpha-specific positive and negative selection. Immunol Rev. 1993; 131:131-168. (Biology). View Reference
  4. Tomonari K. Negative selection of Tcra-V8+CD8+ T cells by MHC class I molecules. Immunogenetics. 1992; 35(5):291-295. (Biology). View Reference
View All (4) View Less
553377 Rev. 10

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