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Flow cytometric analysis of T- and B- Cell Activation Antigen on activated Mouse lymphocytes. Concanavalin A (Con A)-stimulated (3 days) Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either BD Horizon™ BV421 Rat IgM, κ Isotype Control (Cat No. 562708; dashed line histogram) or with the BD Horizon™ BV421 Rat Anti-Mouse T- and B- Cell Activation Antigen antibody (Cat No. 568851/568852; solid line histogram) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histogram showing Mouse T- and B- Cell Activation Antigen expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphoblasts. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Flow Cytometer System and FlowJo™ software.
BD Horizon™ BV421 Rat Anti-Mouse T- and B-Cell Activation Antigen
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
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- An isotype control should be used at the same concentration as the antibody of interest.
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Companion Products
The GL7 antibody specifically recognizes the T- and B- Cell Activation Antigen which is also known as, the GL7 antigen. The GL7 antigen is a 35-kDa cell-surface protein that is expressed on T and B lymphocytes activated in vitro, on bone marrow Pre-B-II cells, germinal-center B cells, and the subpopulation of thymocytes that coexpress high CD3e levels. The GL7 antibody recognizes an epitope containing nonsulfated α2-6-sialyl-LacNAc. There is strain variability with respect to GL7 antigen distribution on thymocytes and Con A-activated spleen cells. GL7 antigen expression is found to be higher on BALB/c mouse leucocytes than on C57BL/6 mouse counterparts. The GL7 antibody reportedly may crossreact with epitopes on molecules expressed by certain rat and human leucocyte subsets.
Development References (9)
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Balogh A, Adori M, Torok K, Matko J, Laszlo G. A closer look into the GL7 antigen: its spatio-temporally selective differential expression and localization in lymphoid cells and organs in human. Immunol Lett. 2010; 130(1-2):89-96. (Clone-specific: Flow cytometry). View Reference
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Escolano A, Gristick HB, Abernathy ME, et al. Immunization expands B cells specific to HIV-1 V3 glycan in mice and macaques.. Nature. 2019; 570(7762):468-473. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Han S, Dillon SR, Zheng B, Shimoda M, Schlissel MS, Kelsoe G. V(D)J recombinase activity in a subset of germinal center B lymphocytes. Science. 1997; 278(5336):301-305. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Han S, Zheng B, Schatz DG, Spanopoulou E, Kelsoe G. Neoteny in lymphocytes: Rag1 and Rag2 expression in germinal center B cells. Science. 1996; 274(5295):2094-2097. (Clone-specific: Flow cytometry). View Reference
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Han S, Zheng B, Takahashi Y, Kelsoe G. Distinctive characteristics of germinal center B cells. Semin Immunol. 1997; 9(4):255-260. (Clone-specific). View Reference
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Hathcock KS, Pucillo CE, Laszlo G, Lai L, Hodes RJ. Analysis of thymic subpopulations expressing the activation antigen GL7. Expression, genetics, and function. J Immunol. 1995; 155(10):4575-4581. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Hatzi K, Geng H, Doane AS, et al. Histone demethylase LSD1 is required for germinal center formation and BCL6-driven lymphomagenesis.. Nat Immunol. 2019; 20(1):86-96. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Laszlo G, Hathcock KS, Dickler HB, Hodes RJ. Characterization of a novel cell-surface molecule expressed on subpopulations of activated T and B cells. J Immunol. 1993; 150(12):5252-5262. (Immunogen: Immunoprecipitation). View Reference
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Naito Y, Takematsu H, Koyama S, et al. Germinal center marker GL7 probes activation-dependent repression of N-glycolylneuraminic acid, a sialic acid species involved in the negative modulation of B-cell activation.. Mol Cell Biol. 2007; 27(8):3008-22. (Clone-specific: Flow cytometry). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.