Anti t bet

Manufactured by BD

Sourced in France

Anti-T-bet is a laboratory reagent used for the detection and quantification of the T-bet transcription factor. T-bet is a key regulator of Th1 cell differentiation and is essential for the development of cell-mediated immunity. Anti-T-bet can be utilized in various immunological assays and research applications.

Automatically generated - may contain errors

Lab products found in correlation

Anti rorγt, by BD (3 mentions) Anti gata3, by BD (2 mentions) Cytofix cytoperm kit, by BD (2 mentions) Golgistop, by BD (2 mentions) Red blood cell lysis buffer, by Merck Group (1 mentions) Anti il 10, by BD (1 mentions) Fc block anti cd16 32, by Thermo Fisher Scientific (1 mentions) Fixation and permeabilization solution, by Thermo Fisher Scientific (1 mentions) Live dead, by Thermo Fisher Scientific (1 mentions) Anti rorγt q31 378, by BD (1 mentions) Brefeldin a, by Merck Group (1 mentions) Ionomycin, by Merck Group (1 mentions) Software, by FlowJo (1 mentions) Facscelesta, by BD (1 mentions) Facscanto flow cytometer, by BD (1 mentions) Bcl 6, by BD (1 mentions) Recombinant human il 12, by R&D Systems (1 mentions) Il 23, by Thermo Fisher Scientific (1 mentions) Anti pstat3, by BD (1 mentions) Percp cy5.5 anti ifn γ, by Thermo Fisher Scientific (1 mentions)

7 protocols using anti t bet

Immunization and T-cell Response Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

For protein immunization, C57BL/6 mice were immunized with Ag85B or MAP1889c (50 μg/mice) via intravenous injection administered three times at 2-week intervals. Two weeks after the final immunization, the spleens were isolated from the mice. The single-cell suspensions were then filtered through a 40-μm cell nylon mesh cell strainer, treated with red blood cell lysis buffer (Sigma) for 5 min, and washed twice with RPMI 1640 medium supplemented with 2% FBS. Single-cell suspensions of the spleen of immunized mice were stimulated with Ag85B (10 μg/mL) or MAP1889c (10 μg/mL) for 12 h at 37 °C in the presence of GolgiStop (BD Biosciences). The cells were first blocked with Fc Block (anti-CD16/32; eBioscience) for 15 min at 4 °C and then stained with fluorochrome-conjugated anti-CD4 or anti-CD8 Ab (BD Biosciences) for 30 min at 4 °C. Cells stained with the appropriate isotype-matched immunoglobulins were used as a negative control. The cells were fixed and permeabilized using a Cytofix/Cytoperm kit (BD Biosciences) according to the manufacturer’s instructions. Intracellular anti-IL-10, anti-T-bet, or anti-GATA-3 (BD Biosciences) levels were detected with fluorescein-conjugated antibodies in permeation buffer. Then, the samples were detected by flow cytometry.

Park H.S., Back Y.W., Son Y.J, & Kim H.J. (2020). Mycobacterium avium subsp. paratuberculosis MAP1889c Protein Induces Maturation of Dendritic Cells and Drives Th2-Biased Immune Responses. Cells, 9(4), 944.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

The cells were preincubated for 20 min with anti-CD16/32 (2.4G2, Fc block from BioXCell) and stained for viability with LIVE/DEAD^® (Invitrogen, Carlsbad, CA) for 20 min on ice. Cells were surface stained for flow cytometry with the combination of the following antibodies (eBioscience or Biolegend): Anti-CD45 (104), anti-CD3 (145-2C11), anti-Thy1.2 (30-H12), anti-B220 (RA3-6B2), anti-Ter-119 (TER119), anti-Gr-1 (RB6-8C5), anti-CD11c (N418), anti-CD5 (53-7.3), anti-Eomes (Dan11mag), anti-T-bet (4B10), anti-RORγt (Q31-378, BD Pharmingen), anti-IFNγ (XMG1.2), anti-IL-17a (TC11-18H10.1), anti-Ly6G (1A8), anti-CD11b (M1/70), anti-CD64 (X54-5/7.1), anti-CD103 (2E7), anti-MHCII (M5/114.15.2). For intracellular cytokine staining was cells (1 x 10⁶ cells) were restimulated for 4 hours at 37°C with PMA (50 ng/ml) and ionomycin (500 nM) in the presence of brefeldin A (5 μg/ml) (all from Sigma-Aldrich, St. Louis MO) and fixed and permeabilized using fixation and permeabilization solution (eBiosciences). All cells were analyzed on either a FACS Celesta or LSR II (BD Biosciences) and analyzed with FlowJo software (FlowJo LLC).

Jing X., Korchagina A.A., Shein S.A., Muraoka W.T., Koroleva E, & Tumanov A.V. (2021). IL-23 Contributes to Campylobacter jejuni-Induced Intestinal Pathology via Promoting IL-17 and IFNγ Responses by Innate Lymphoid Cells. Frontiers in Immunology, 11, 579615.

+ Open protocol

+ Expand

T Cell Differentiation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Naïve CD4⁺CD25^- T cells were differentiated to the different effector subsets as detailed in Supplemental Experimental Procedures. For proliferation assays, cells were labeled Cell Violet (1 μM) and activated with anti-CD3/CD28 or differentiated as indicated. For intracellular staining of transcription factors, cells were differentiated during 3 days, collected with 5 mM EDTA in PBS, fixed, permeabilized and stained with anti-RORγT and anti-T-bet (BD biosciences). For intracellular cytokine staining, cells were restimulated for 4 hr with PMA and ionomycin in the presence of brefeldin, permeabilized with 0.5% saponin and stained with anti-CD4, anti-IFN-γ, anti-IL-17 and anti-Foxp3. Data were acquired on a FACSCanto flow cytometer (BD Bioscience) and analyzed using the FLOWJO software (Tree Star).

Baixauli F., Acín-Pérez R., Villarroya-Beltrí C., Mazzeo C., Nuñez-Andrade N., Gabandé-Rodriguez E., Dolores Ledesma M., Blázquez A., Martin M.A., Falcón-Pérez J.M., Redondo J.M., Enríquez J.A, & Mittelbrunn M. (2015). Mitochondrial respiration controls lysosomal function during inflammatory T cell responses. Cell metabolism, 22(3), 485-498.

+ Open protocol

+ Expand

Multiparametric Profiling of T Cell Subsets

Check if the same lab product or an alternative is used in the 5 most similar protocols

eFluor660-anti–IL-21, Alexa Fluor 488–anti–IL-10, PerCP-Cy5.5-anti–IFN-γ, FITC-anti-CD45RA, PE-anti-ICOS, and anti-Tbet, and biotin–PD-1 were from eBioscience. Alexa Fluor 488–anti-GATA3, Alexa Fluor 647–anti-CXCR5, anti-pSTAT4, anti-pSTAT5, anti-pSTAT6, APC-anti-CD10, APC-Cy7-anti-CD4, BV605-anti-IgG, BV421-CD40L, BV711-anti–IL-2, PE-anti-pSTAT1, anti-RORγt, and Bcl-6, PE-Cy7-anti-CD25, and anti-CD27, PerCP-Cy5.5-anti-CD127, anti-pSTAT3, and anti-Tbet, SA-PerCpCy5.5, and IFN-γ were obtained from BD. Pacific Blue–anti-CD20 and SA-BV605 were purchased from BioLegend. Recombinant human IL-12 was purchased from R&D Systems. TGF-β, IL-1β, IL-6, IL-21, and IL-23 were obtained from PeproTech. PGE2 was purchased from Sigma-Aldrich. Human IL-4 and IL-10 were provided by R. de Waal Malefyt (DNAX Research Institute, Palo Alto, CA).

Ma C.S., Wong N., Rao G., Nguyen A., Avery D.T., Payne K., Torpy J., O’Young P., Deenick E., Bustamante J., Puel A., Okada S., Kobayashi M., Martinez-Barricarte R., Elliott M., Sebnem Kilic S., El Baghdadi J., Minegishi Y., Bousfiha A., Robertson N., Hambleton S., Arkwright P.D., French M., Blincoe A.K., Hsu P., Campbell D.E., Stormon M.O., Wong M., Adelstein S., Fulcher D.A., Cook M.C., Stepensky P., Boztug K., Beier R., Ikincioğullari A., Ziegler J.B., Gray P., Picard C., Boisson-Dupuis S., Phan T.G., Grimbacher B., Warnatz K., Holland S.M., Uzel G., Casanova J.L, & Tangye S.G. (2016). Unique and shared signaling pathways cooperate to regulate the differentiation of human CD4+ T cells into distinct effector subsets. The Journal of Experimental Medicine, 213(8), 1589-1608.

+ Open protocol

+ Expand

Epigenetic Profiling by ChIP-seq and RNA-seq

Check if the same lab product or an alternative is used in the 5 most similar protocols

ChIP-seq assays were performed as described previously ^{3 ,18}. In brief, cells for ChIP-seq were fixed for 10 minutes in 1% formaldehyde and sonicated and chromatin immunoprecipitation was performed with anti-H3K4me1(ab8895, Abcam), anti-H3K4me2 (ab32356, Abcam), anti-H3K4me3 (17–614, Millipore), anti-H3K9me3 (ab8898, Abcam), anti-H3K27ac (ab4729, Abcam), anti-H3K27me3 (07–449, Millipore), anti-MLL4 ^{19 ,20}, anti-GATA3 (558686, BD bioscience), anti-T-bet (561263, BD bioscience). DNA was end-repaired using an End-It DNA-Repair kit (Epicentre), and was indexed, amplified and sequenced on an Illumina HiSeq-2500. RNA-seq libraries using poly-A RNA isolated from both the WT and MLL4 KO cells were prepared as previous described ²¹ and sequenced by Illumina HiSeq-2500.

Cui K., Chen Z., Cao Y., Liu S., Ren G., Hu G., Fang D., Wei D., Liu C., Zhu J., Wu C, & Zhao K. (2023). Restraint of IFN-γ expression through a distal silencer CNS–28 for tissue homeostasis. Immunity, 56(5), 944-958.e6.

+ Open protocol

+ Expand

Cytokine and Transcription Factor Analysis in Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For measurement of intracellular cytokine expression, cells were isolated ex vivo and stimulated with 50 ng/ml PMA and 500 ng/ml ionomycin, in the presence of GolgiStop (BD Biosciences) for 3 h. Dead cells were excluded by Fixable Viability Dye eFluor 450 (eBioscience). Cells were stained with antibodies to surface antigens, and then fixed and permeabilized with the Cytofix/Cytoperm kit (BD Biosciences), followed by staining with anti-IL-5 (Biolegend) and anti-IL-13 (eBioscience).
For analysis of transcription factor expression, cells were stained with antibodies to surface antigens, then fixed and permeabilized with the Foxp3/Transcription Factor Staining Buffer Set (eBioscience) according to the manufacturer’s instructions, and stained with anti-GATA3 (eBioscience), anti-RORγt, anti-T-bet (BD Biosciences), or anti-Ki-67 (eBioscience).
For measurement of intracellular HIF1α, Lin⁻ bone marrow cells were cultured in complete media with 10 ng/ml IL-7 (Peprotech) and 10 ng/ml IL-33 (R&D Systems) for 4 days. Cells were collected and stained with antibodies to surface antigens, fixed and permeabilized with the Foxp3/Transcription Factor Staining Buffer Set (eBioscience) according to the manufacturer’s instructions, and then stained with anti-HIF1α (R&D Systems).

Li Q., Li D., Zhang X., Wan Q., Zhang W., Zheng M., Zou L., Elly C., Lee J.H, & Liu Y.C. (2018). The E3 ligase VHL promotes group 2 innate lymphoid cell maturation and function via inhibition of glycolysis and induction of interleukin 33 receptor. Immunity, 48(2), 258-270.e5.

+ Open protocol

+ Expand

Multiparametric Analysis of MAIT Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were stained with fixable viability dye (eBioscience-Thermo Fisher Scientific, Bleiswijk, The Netherlands) and then with the following antibodies for surface staining: anti-TCRVα7.2, anti-CD161, anti-CD3, anti-ST2, anti-CD4 and anti-CD8 (SONY biotechnology, Weybridge, United Kingdom) (eBiosciences-Thermo Fisher Scientific) (Biolegend, Amsterdam, The Netherlands). MAIT cells were identified as TCRVα7.2⁺CD161⁺ or MR1 5-OP-RU tetramer⁺ cells. MR1 6-FP tetramer⁺ was used as negative control (NIH Tetramer Facility, Atlanta, GA, USA) (Figure S1). For intracellular cytokine staining, cells were fixed in 4% paraformaldehyde, washed, and permeabilized with 0.5% saponin (Sigma-Aldrich), and then incubated with anti-IFNγ, anti-TNFα, anti-Granzyme B or isotype controls (eBiosciences-Thermo Fisher Scientific, SONY biotechnology or BioLegend). In some experiments, MAIT cells were stained intracellularly with anti-Tbet (BD Biosciences, Grenoble, France) and anti-PLZF (R&D Systems) antibodies using a transcription factor buffer set (BD Biosciences). Events were acquired on a FACS LSR Fortessa (BD Biosciences) and analyzed using FlowJo software v10.7.1 (Becton, Dickinson and Company, Ashland, OR, USA).

Azzout M., Dietrich C., Machavoine F., Gastineau P., Bottier A., Lezmi G, & Leite-de-Moraes M. (2021). IL-33 Enhances IFNγ and TNFα Production by Human MAIT Cells: A New Pro-Th1 Effect of IL-33. International Journal of Molecular Sciences, 22(19), 10602.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!