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PE Mouse Anti-Human Cβ1 TCR
PE Mouse Anti-Human Cβ1 TCR
Multicolor flow cytometric analysis of TCR Cβ1 expression by human peripheral blood lymphocytes. Human peripheral blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the cells were stained with either APC Mouse Anti-Human CD3 (Left Plots; Cat. No. 555335/561810/561811), BD Horizon™ BV421 Mouse Anti-Human CD4 (Middle Plots; Cat. No. 562424/562425), or APC Mouse Anti-Human CD8 (Right Plots; Cat. No. 555369/561421/561952/561953) antibody and either PE Mouse IgG2a, κ Isotype Control (Cat. No. 554648; Top Plots) or PE Mouse Anti-Human TCR Cβ1 antibody (Cat. No. 565776; Bottom Plots) at 1 µg/test. Two-color flow cytometric contour plots showing the correlated expression of CD3, CD4 or CD8 versus TCR Cβ1 (or Ig Isotype Control staining) were derived from gated events with the forward and side light-scatter characteristics of viable human peripheral blood lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.
Multicolor flow cytometric analysis of TCR Cβ1 expression by human peripheral blood lymphocytes. Human peripheral blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. After washing, the cells were stained with either APC Mouse Anti-Human CD3 (Left Plots; Cat. No. 555335/561810/561811), BD Horizon™ BV421 Mouse Anti-Human CD4 (Middle Plots; Cat. No. 562424/562425), or APC Mouse Anti-Human CD8 (Right Plots; Cat. No. 555369/561421/561952/561953) antibody and either PE Mouse IgG2a, κ Isotype Control (Cat. No. 554648; Top Plots) or PE Mouse Anti-Human TCR Cβ1 antibody (Cat. No. 565776; Bottom Plots) at 1 µg/test. Two-color flow cytometric contour plots showing the correlated expression of CD3, CD4 or CD8 versus TCR Cβ1 (or Ig Isotype Control staining) were derived from gated events with the forward and side light-scatter characteristics of viable human peripheral blood lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
T-cell receptor beta locus; TCRB; TRB; TRB@; TRCB1
Human (QC Testing)
Mouse IgG2a, κ
Lymphoid cells from Mouse transgenic for Human HA1.7 cell TCRbeta
Flow cytometry (Reported)
5 µl/test
AB_2739349
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 to the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565776 Rev. 2
Antibody Details
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JOVI.1

The JOVI.1 monoclonal antibody recognizes an epitope common to a large proportion of human CD4+ or CD8+ T lymphocytes that express the T cell receptor beta chain (TCRβ). The antibody was generated from a mouse immunized with transgenic mouse lymphoid cells that expressed the rearranged human Vβ3-Cβ1 TCR chain derived from the cloned human HA1.7 T helper cell. This antibody reacts with TCR-Cβ1+ T cells and one of several different TCRβ V regions, but not with TCR-Cβ2+ T cells. JOV1.1 antibody reportedly recognized several Cβ1 TCR expressing cell lines or clones including Jurkat, CH7C17, and HA1.7 cells. The JOVI.1 antibody can be used to stimulate proliferative responses by JOVI.1-positive T cells. It can also reportedly be used for immunoprecipitation and to stain JOVI.1+ T cells in frozen tissue sections.

        

565776 Rev. 2
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.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
565776 Rev.2
Citations & References
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Development References (3)

  1. Gil D, Schamel WWA, Montoya M, Sanchez-Madrid F, and Alarcon B. Recruitment of Nck by CD3-epsilon reveals a ligand-induced conformational change essential for T cell receptor signaling and synapse formation. Cell. 2002; 109(7):901-912. (Clone-specific: Activation, Functional assay, Immunoprecipitation). View Reference
  2. San José E, Alarcón B. Receptor engagement transiently diverts the T cell receptor heterodimer from a constitutive degradation pathway.. J Biol Chem. 1999; 274(47):33740-6. (Clone-specific: Immunoprecipitation). View Reference
  3. Viney JL, Prosser HM, Hewitt CR, Lamb JR, Owen MJ. Generation of monoclonal antibodies against a human T cell receptor beta chain expressed in transgenic mice. Hybridoma. 1992; 11(6):701-713. (Immunogen: Activation, Bioassay, Flow cytometry, Functional assay, Immunohistochemistry, Immunoprecipitation, Radioimmunoassay, Stimulation). View Reference
565776 Rev. 2

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


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.