Multicolor flow cytometric analysis of TCR βF1 expression by human peripheral blood lymphocytes. Whole blood was surface stained with FITC Mouse Anti-Human TCR αβ (Cat. No. 555547; Top Plots) and PE Mouse Anti-Human TCR γδ (Cat. No. 561994; Bottom Plots) antibodies. Erythrocytes were lysed and leucocytes were fixed with 1x BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049). After permeabilization with BD™ Phosflow Perm Buffer I (Cat. No. 557885), the cells were then stained with either APC Mouse IgG1, κ Isotype Control (Cat. No. 554681; Left Plots) or APC Mouse Anti-Human TCR βF1 antibody (Cat. No. 566052; Right Plots). Two color contour plots showing the correlated expression of TCR βF1 (or Ig Isotype control staining) versus TCR αβ or TCR γδ were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
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Multicolor flow cytometric analysis of TCR βF1 expression by human peripheral blood lymphocytes. Whole blood was surface stained with FITC Mouse Anti-Human TCR αβ (Cat. No. 555547; Top Plots) and PE Mouse Anti-Human TCR γδ (Cat. No. 561994; Bottom Plots) antibodies. Erythrocytes were lysed and leucocytes were fixed with 1x BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049). After permeabilization with BD™ Phosflow Perm Buffer I (Cat. No. 557885), the cells were then stained with either APC Mouse IgG1, κ Isotype Control (Cat. No. 554681; Left Plots) or APC Mouse Anti-Human TCR βF1 antibody (Cat. No. 566052; Right Plots). Two color contour plots showing the correlated expression of TCR βF1 (or Ig Isotype control staining) versus TCR αβ or TCR γδ were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
TCRb; TCRB; TCR beta; TCR-beta; TRB@; TCR Beta F1; TCR BF1; TCR Cβ
Human (QC Testing)
Mouse BALB/c IgG1, κ
Purified Human αβ TCR Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2739482
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 APC under optimum conditions, and unconjugated antibody and free APC were removed.
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lyse/Fix Buffer 5X RUO
Size
250 mL
Cat No.
558049
Perm/Wash Buffer I RUO
Size
125 mL
Cat No.
557885
APC Mouse IgG1 κ Isotype Control RUO
Size
0.1 mg
Cat No.
554681
FITC Mouse Anti-Human TCR αβ RUO
Size
100 Tests
Cat No.
555547
566052 Rev. 2
Antibody Details
8A3 (βF1)
The 8A3 monoclonal antibody is also known as the β Framework 1 (βF1) antibody. After treatment of lymphoid cells with permeabilization reagents, this antibody recognizes a common epitope on the β chain of the T Cell Receptor (TCR) for antigen which is expressed by thymocytes and peripheral T lymphocytes. It does not react with γδ TCR-bearing T cells. The 8A3 antibody does not stain the surface of living cells. This suggests that the TCR β constant region epitope recognized by the 8A3 antibody is normally hidden on viable cells.
566052 Rev. 2
Format Details
APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
APC
Red 627-640 nm
651 nm
660 nm
566052 Rev.2
Citations & References
View product citations for antibody "566052" on CiteAb
Development References (5)
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Asnafi V, Beldjord K, Boulanger E, et al. Analysis of TCR, pT alpha, and RAG-1 in T-acute lymphoblastic leukemias improves understanding of early human T-lymphoid lineage commitment. Blood. 2003; 101(7):2693-2703. (Clone-specific: Flow cytometry). View Reference
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Brenner MB, McLean J, Scheft H, Warnke RA, Jones N, Strominger JL. Characterization and expression of the human alpha beta T cell receptor by using a framework monoclonal antibody. J Immunol. 1987; 138(5):1502-1509. (Immunogen: Flow cytometry, Fluorescence microscopy, Immunohistochemistry, Immunoprecipitation, Radioimmunoassay, Western blot). View Reference
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Fraser JD, Strominger JL. A solubilized T-cell receptor from a human leukemia cell line binds to a ligand in the absence of MHC products.. Immunogenetics. 1988; 28(2):108-16. (Clone-specific: Functional assay). View Reference
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Ramiro AR, Trigueros C, Márquez C, San Millán JL, Toribio ML. Regulation of pre-T cell receptor (pT alpha-TCR beta) gene expression during human thymic development. J Exp Med. 1996; 184(2):519-530. (Clone-specific: Flow cytometry). View Reference
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Torres PS, Alcover A, Zapata DA, et al. TCR dynamics in human mature T lymphocytes lacking CD3 gamma. J Immunol. 2003; 170(12):5947-5955. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
566052 Rev. 2
Please refer to Support Documents for Quality Certificates
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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.