Bcl 6

Manufactured by BioLegend

Sourced in United States

Bcl-6 is a laboratory reagent used in flow cytometry applications. It functions as a transcriptional repressor involved in the regulation of B-cell differentiation.

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Lab products found in correlation

Ifn γ, by BioLegend (2 mentions) Cxcr5, by BioLegend (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Lsrfortessa flow cytometer, by FlowJo (1 mentions) Il 17a, by BioLegend (1 mentions) Anti biotin, by BioLegend (1 mentions) Igg2a, by BioLegend (1 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) Phospho ikkα β, by Cell Signaling Technology (1 mentions) Il 21, by Abcam (1 mentions) P100 52, by Cell Signaling Technology (1 mentions) Phospho stat3, by Cell Signaling Technology (1 mentions) Stat3, by Abcam (1 mentions) Live dead fixable blue, by Thermo Fisher Scientific (1 mentions) Tcf 1, by BD (1 mentions) Tim 3, by BioLegend (1 mentions) Intracellular fixation permeabilization buffer set, by Thermo Fisher Scientific (1 mentions) Anti cd3 ab, by BD (1 mentions) Brefeldin a, by Thermo Fisher Scientific (1 mentions) Anti cd28 ab, by BD (1 mentions)

5 protocols using bcl 6

Flow Cytometry Analysis of Immune Cells

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Fluorophor-conjugated antibodies against the indicated surface markers were used to stain RBC-depleted splenocytes, LN cells, BM cells and peripheral blood cells. For intracellular staining, cells were first surface stained for lineage markers, fixed, permeabilized and stained with antibodies against Bcl6, IL-17A, IFNγ, or Ki67 (Biolegend). Stained cells were analyzed using a BD LSR Fortessa flow cytometer, and further data analysis was carried out using FlowJo software.

Hu R., Kagele D.A., Huffaker T.B., Runtsch M.C., Alexander M., Liu J., Bake E., Su W., Williams M.A., Rao D.S., Möller T., Garden G.A., Round J.L, & O’Connell R.M. (2014). miR-155 promotes T follicular helper cell accumulation during chronic, low-grade inflammation.. Immunity, 41(4), 605-619.

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Multicolor Flow Cytometry for Mouse Lymphoid Tissue Analysis

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For mouse T cell and B cell analysis in lymphoid tissues, LN cells were stained for viability and extracellular antigens with directly labeled antibodies, CD95 (BioLegend 152606), IgD (BioLegend 405710), GL7 (BioLegend 144617), live/dead fixable yellow dead cell staining (Thermo Fisher Scientific L34968), CD19 (BioLegend 115555), biotinylated SARS-CoV-2-S1 protein (BioLegend 793804), anti-biotin (BioLegend 409003), CD3 (BioLegend 100220), CD3 (BioLegend 100204), CD4 (BioLegend 100540), ICOS (BioLegend 117420), CD8 (BioLegend 557654), CD45 (BioLegend 103134), CXCR5 (BioLegend 145504), PD-1 (BioLegend 135231), CD38 (BioLegend 102712), IgG1 (BioLegend 406620), and IgG2a (BioLegend 407114). Cells were washed, fixed, and permeabilized by using the Ebioscience Foxp3/Transcription Factor Staining Buffer Set (Thermo Scientific 00-5523-00) kit according to manufacture instructions. Permeabilized cells were intracellularly stained with Bcl-6 (BioLegend 358512). Cells were acquired on an Attune NxT Flow Cytometer and analyzed with Attune NxT software v.4.2.0.

Hu Q., Zhao Y., Shaabani N., Lyu X., Powers C., Sun H., Cruz V., Stegman K., Xu J., Fossier A., Huang Y., Ho G., Kao Y., Wang Z., Wang Z., Hu Y., Zheng Y., Kyaw L., Zuluaga C., Wang H., Pei H., Allen R., Xie H., Ji H, & Chen R. (2022). Chimeric mRNA-based COVID-19 vaccine induces protective immunity against Omicron and Delta variants. Molecular Therapy. Nucleic Acids, 30, 465-476.

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Western Blotting Technique for Protein Analysis

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Western blotting was performed as previously described (27 (link)). Briefly, proteins were extracted using RIPA buffer (Gensharebio, Xi’an, China). Protein concentration was measured with a Pierce BCA Protein Assay kit (Thermo Fisher Scientific, Waltham, MA, USA). Equal quality of protein was loaded in 10% SDS-PAGE gel, transferred onto nitrocellulose membranes and probed with primary antibodies. The primary antibodies employed were Bcl-6 (BioLegend), CXCR5 (EMD Millipore, Billerica, MA, USA), IL-21 (Abcam, Cambridge, UK), STAT3, Phospho-STAT3, iKK-α, Phospho-iKKα/β, NF-κB-inducing kinase (NIK), P100/52, and B lymphocyte-induced maturation protein-1 (Blimp-1) (all from Cell Signaling Technology, Danvers, MA, USA).

Guo J., Zhao C., Wu F., Tao L., Zhang C., Zhao D., Yang S., Jiang D., Wang J., Sun Y., Li Z., Li H, & Yang K. (2018). T Follicular Helper-Like Cells Are Involved in the Pathogenesis of Experimental Autoimmune Encephalomyelitis. Frontiers in Immunology, 9, 944.

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Multiparametric Phenotyping of T-cells

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LIVE/DEAD fixable blue (ThermoFisher, Cat #: L23962), Rat anti-mouse T-BET (Biolegend, 644806, 1:100), BLIMP1 (Biolegend, Cat #: 150008, 1:100), PD-1 (Biolegend, Cat #: 135219, 1:100), BCL6 (Biolegend, Cat #: 358512, 1:200), CD8 (Biolegend, Cat #: 100766, 1:200), CD44 (Biolegend, Cat #: 103057, 1:500), IFN-γ (Biolegend, Cat #: 505838, 1:400), TIM-3 (Biolegend, Cat #: 134013, 1:100) and TCF1 (BD Pharmingen, Cat #: 564217, 1:200) were utilized.

Marro B.S., Zak J., Zavareh R.B., Teijaro J.R., Lairson L.L, & Oldstone M.B. (2019). Discovery of small molecules for the reversal of T cell exhaustion. Cell reports, 29(10), 3293-3302.e3.

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Intracellular Staining and Flow Cytometry

Cited 1 time

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The procedure of cell staining and flow cytometry was previously described (5 (link)). For intracellular staining of IL-21, gastric CD4⁺T cells were restimulated with 1 μg/ml anti-CD3 Ab (Clone UCHT1, BD), 1 μg/ml anti-CD28 Ab (Clone CD28.2, BD), and 3 μg/ml brefeldinA (eBioscience, CA, USA) for 12 h. The intracellular fixation/permeabilization buffer set (eBioscience, CA, USA) was used in IL-21 staining. Flow cytometric detection was performed with FACS Canto II (BD, NJ, USA). Data was analyzed by the FlowJo software (Tree Star). The anti-human antibodies used in this study were purchased from Biolegend, BD Biosciences, or Miltenyi Biotec, namely: CD3 (Clone UCHT1, BioLegend), CD4 (Clone OKT4, BioLegend), GITR (Clone 108-17, BioLegend), GITRL (Clone REA841, Miltenyi), IL-21 (Clone 3A3-N2.1, BD), BCL6 (Clone 7D1, BioLegend), STAT3 Phospho (Ser727) (Clone A16089B, BioLegend), BTLA (Clone MIH26, BioLegend), CXCR5 (Clone J252D4, BioLegend), and PD-1 (Clone EH12.2H7, BioLegend).

Ming S., Yin H., Li X., Gong S., Zhang G, & Wu Y. (2021). GITR Promotes the Polarization of TFH-Like Cells in Helicobacter pylori-Positive Gastritis. Frontiers in Immunology, 12, 736269.

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