BD Horizon™ RB744 Mouse Anti-Human TCR γδ
Clone 11F2 (RUO)
Two-color flow cytometric analysis of TCR γδ expression on Human peripheral blood T lymphocytes. Human whole blood was stained with BD Horizon™ BUV395 Mouse Anti-Human CD3 antibody (Cat. No. 563546) and with either BD Horizon™ RB744 Mouse IgG1, κ Isotype Control (Cat No. 570519; Left Plot) or BD Horizon™ RB744 Mouse Anti-Human TCR γδ antibody (Cat No. 570494/570495; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing the correlated express of TCR γδ (or Ig Isotype control staining) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software.
Two-color flow cytometric analysis of TCR γδ expression on Human peripheral blood T lymphocytes. Human whole blood was stained with BD Horizon™ BUV395 Mouse Anti-Human CD3 antibody (Cat. No. 563546) and with either BD Horizon™ RB744 Mouse IgG1, κ Isotype Control (Cat No. 570519; Left Plot) or BD Horizon™ RB744 Mouse Anti-Human TCR γδ antibody (Cat No. 570494/570495; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing the correlated express of TCR γδ (or Ig Isotype control staining) versus CD3 was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software.
Product Details
BD Horizon™
TCRgd; γδ TCR; TRG@/TRD@; TCRG/TCRD; TCR gamma delta
Human (QC Testing)
Mouse BALB/c IgG1
Sepharose® bead/CD3/γ/δ TCR complex
Flow cytometry (Routinely Tested)
5 µl/test
6964, 6965
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 unreacted dye was 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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
BUV395 Mouse Anti-Human CD3 RUO
Size
100 Tests
Cat No.
563546
RB744 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
570519
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
570495 Rev. 1
Antibody Details
11F2
The 11F2 monoclonal antibody specifically reacts with a framework epitope of the γ/δ TCR. The γ/δ TCR is a heterodimeric glycoprotein that is noncovalently associated with the CD3 antigen complex. The γ and δ TCR chains are composed of constant and variable regions, each encoded by distinct gene segments. The γ chain forms either disulfide-linked or non-disulfide-linked heterodimers with the δ-subunit. TCR γ/δ is present on a minor subset of T lymphocytes in peripheral blood, thymus, spleen, and lymph node. TCR γ/δ-positive T lymphocytes comprise 1% to 9% of normal peripheral blood lymphocytes and less than 2% of normal thymocytes. The 11F2 antibody is mitogenic for γ/δ-TCR-bearing T lymphocytes.
570495 Rev. 1
Format Details
RB744
The BD Horizon RealBlue™ 744 (RB744) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 746-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB744 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), we recommend using an optical filter centered near 750-nm (e.g., a 750/60-nm bandpass filter).
RB744
Blue 488 nm
498 nm
746 nm
570495 Rev.1
Citations & References
View product citations for antibody "570495" on CiteAb
Development References (6)
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Borst J, van Dongen JJ, Bolhuis RL, et al. Distinct molecular forms of human T cell receptor gamma/delta detected on viable T cells by a monoclonal antibody.. J Exp Med. 1988; 167(5):1625-44. (Immunogen: Electron microscopy, ELISA, Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
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Hertoghs N, Schwedhelm KV, Stuart KD, McElrath MJ, De Rosa SC. OMIP-064: A 27-Color Flow Cytometry Panel to Detect and Characterize Human NK Cells and Other Innate Lymphoid Cell Subsets, MAIT Cells, and γδ T Cells.. Cytometry A. 2020; 97(10):1019-1023. (Clone-specific: Flow cytometry). View Reference
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Ichinohasama R, Miura I, Takahashi T, et al. Peripheral CD4+ CD8- gammadelta T cell lymphoma: a case report with multiparameter analyses.. Hum Pathol. 1996; 27(12):1370-7. (Clone-specific: Flow cytometry). View Reference
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Lanier LL, Federspiel NA, Ruitenberg JJ, et al. The T cell antigen receptor complex expressed on normal peripheral blood CD4-, CD8- T lymphocytes. A CD3-associated disulfide-linked gamma chain heterodimer.. J Exp Med. 1987; 165(4):1076-94. (Clone-specific: Flow cytometry). View Reference
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Lanier LL, Ruitenberg J, Bolhuis RL, Borst J, Phillips JH, Testi R. Structural and serological heterogeneity of gamma/delta T cell antigen receptor expression in thymus and peripheral blood.. Eur J Immunol. 1988; 18(12):1985-92. (Clone-specific: Flow cytometry, Immunoprecipitation). View Reference
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Testi R, Lanier LL. Functional expression of CD28 on T cell antigen receptor gamma/delta-bearing T lymphocytes.. Eur J Immunol. 1989; 19(1):185-8. (Clone-specific: Flow cytometry, Functional assay). View Reference
570495 Rev. 1
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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.