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Anti t bet 4b10

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Anti-T-bet (4B10) is a mouse monoclonal antibody that specifically recognizes the Th1-specific transcription factor T-bet. T-bet is a critical regulator of Th1 cell differentiation and is involved in the production of Th1 cytokines, such as IFN-gamma. This antibody can be used for applications such as Western blotting, immunoprecipitation, and flow cytometry to detect and study T-bet expression in various cell types.

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Lab products found in correlation

Anti ubiquitin, by Merck Group (2 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (2 mentions) Anti human cd8 rpa t8, by BD (2 mentions) Anti t bet ebio4b10, by Thermo Fisher Scientific (2 mentions) Total nedd4 2, by Cell Signaling Technology (2 mentions) Tcf 1 2206, by Cell Signaling Technology (2 mentions) Anti gata 3 twaj, by Thermo Fisher Scientific (2 mentions) Anti junb 210, by Santa Cruz Biotechnology (2 mentions) Anti foxp3 fjk 16s, by Thermo Fisher Scientific (2 mentions) Mg132, by Merck Group (2 mentions) Anti eomes dan11mag, by Thermo Fisher Scientific (2 mentions) Recombinant il 25, by R&D Systems (2 mentions) Anti nk1, by Thermo Fisher Scientific (2 mentions) Anti ter119, by Thermo Fisher Scientific (2 mentions) Il 33, by R&D Systems (2 mentions) Il 23, by Thermo Fisher Scientific (2 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (2 mentions) Interleukin 3 (il 3), by Thermo Fisher Scientific (2 mentions) Interleukin 7 (il 7), by Thermo Fisher Scientific (2 mentions) Anti cd8a, by Thermo Fisher Scientific (2 mentions)

Spelling variants of this product

T-bet (36 citations) Anti-T-bet (17 citations) T-bet (4B10) (11 citations) Anti-T-Bet (clone 4B10) (2 citations)

8 protocols using anti t bet 4b10

1

Immunological Profiling with Flow Cytometry

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The following antibodies for flow cytometry were from BD Biosciences: anti-B220 (RA3-6B2), anti-CD3 (2C11), anti-CD4 (RM4–5), anti-CD8 (Ly-3), anti-CD25 (PC61), anti-CD44 (IM7), anti-CD124 (mIL4R-M1), anti-IFN-γ (XMG1.2), anti-IL-17a (TC11-18H10), and anti-human CD8 (RPA-T8). Anti-CD62L (MEL-14), anti-IL-13 (eBio13A), anti-IL-5 (TRFK5), anti-T-bet (eBio4B10) and anti-Foxp3 (FJK-16s) were from eBioscience.
The following antibodies for immunoblotting were from Cell Signaling Technologies: Antibody to Rictor (2140), NDRG1 phosphorylated at Thr346 (5482), total NDRG1 (9408), antibody to Akt phosphorylated at S473 (4060), pan-Akt (4685), NEDD4-2 phosphorylated at Ser342 (4080), total NEDD4-2 (4013), TCF-1 (2206), antibody to β-catenin (9562), antibody to phosphorylated β-catenin (9561), antibody to phosphorylated STAT6 (9361). Anti-T-bet (4B10) and anti-GATA-3 (TWAJ) were from eBioscience. Anti-JunB (210) was from Santa Cruz Biotechnology. Anti-ubiquitin was and actin were from Sigma. MG132 was from Sigma, PP242 was from Calbiochem, and CFSE was from Invitrogen.
Heikamp E.B., Patel C.H., Collins S., Waickman A., Oh M.H., Sun I.H., Illei P., Sharma A., Naray-Fejes-Toth A., Fejes-Toth G., Sen J., Horton M.R, & Powell J.D. (2014). The AGC kinase serum- and glucocorticoid-regulated kinase 1 (SGK1) regulates TH1 and TH2 differentiation downstream of mTORC2. Nature immunology, 15(5), 457-464.
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2

Affinity Purification of Tagged Proteins

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HEK293T cells were transiently transfected with expression vectors for mNeonGreen-tagged or HA-tagged proteins using polyethyleneimine. Cells were lysed in lysis buffer (50 mM Tris pH 7.5, 300 mM NaCl, 0.5% IGEPAL CA-630, 1 mM EGTA, 2 mM MgCl2) supplemented with 250 U/ml benzonase (Santa Cruz), 1 mM phenylmethylsulfonyl fluoride, 1 mM DTT and 1X complete EDTA-free protease inhibitor (Roche). The cell lysates were sonicated for 10 sec using a Bioruptor Pico (Diagenode) and then incubated on an over-head shaker for 30 min at 4°C. Insoluble material was removed by centrifugation (17,000xg for 10 min), diluted in an equal volume of lysis buffer without NaCl and incubated with 10 μl mNeonGreen-Trap magnetic agarose beads (ChromoTek) on an over-head shaker for 1 hr at 4°C. Beads were washed 2x with 200 μl lysis buffer containing 150 mM NaCl and 4x with lysis buffer containing 500 mM NaCl, resuspended in 1x Laemmli buffer and incubated at 95°C for 9 min. Proteins were resolved on a 12% Tris-glycine gel and transferred to a nitrocellulose membrane. Membranes were probed with anti-β-Tubulin (ab6046, Abcam), anti-HA (3F10, Roche), anti-FLAG (M2, Sigma), anti-GATA3 (HG3-31 (Santa Cruz), anti-T-bet (4B10, eBioscience), anti-β-actin (4967S, Cell Signaling Technologies), and anti-mNeonGreen (#53061, Cell Signaling Technologies).
Hertweck A., Vila de Mucha M., Barber P.R., Dagil R., Porter H., Ramos A., Lord G.M, & Jenner R.G. (2022). The TH1 cell lineage-determining transcription factor T-bet suppresses TH2 gene expression by redistributing GATA3 away from TH2 genes. Nucleic Acids Research, 50(8), 4557-4573.
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3

Multiparametric Flow Cytometry for Immune Cell Analysis

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Flow cytometry was performed as described previously (11 (link), 14 (link)). In details, single cells were isolated from mouse BM, spleen, and periphery lymph nodes (pLN). If necessary, BM cells were counted by Fuchs-Rosenthal Counting Chamber. Briefly, to analyze the surface marker, cells were blocked with anti-CD16/CD32 antibodies (eBioscience, San Diego, CA) and then stained with surface marker antibodies and washed by PBS. To analyze the intracellular proteins, after staining with surface markers, cells were fixed and permeabilized with Foxp3/Transcription Factor Staining Buffer Kit following the manufacturer's protocols (eBioscience, San Diego, CA).
Antibodies used were as followings: anti-Foxo1 (C29H4) was from Cell Signaling Technology (Boston, MA); anti-NKp46 (29A1.4), anti-CD19 (6D5), anti-CD3 (17A2), anti-Gr1 (RB6-8C5), anti-Ter119 (Ter119), anti-CD4 (GK1.5), anti-CD8 (53-6.7), anti-c-Kit (2B8), anti-Sca-1 (D7), anti-CD127 (A7R34), and anti-CD49b (DX5) were from Biolegend (San Diego, CA); anti-CD27 (LG.3A10), anti-CD11b (M1/70), anti-B220 (RA3-6B2), and anti-CD122 (TM-β1) were form BD Biosciences (San Diego, CA); anti-CD43 (S7), anti-KLRG1 (2F1), anti-CD135(A2F10), anti-Ki-67 (SolA15), and anti-T-bet (4B10) were from eBioscience (San Diego, CA).
Huang P., Wang F., Yang Y., Lai W., Meng M., Wu S., Peng H., Wang L., Zhan R., Imani S., Yu J., Chen B., Li X, & Deng Y. (2019). Hematopoietic-Specific Deletion of Foxo1 Promotes NK Cell Specification and Proliferation. Frontiers in Immunology, 10, 1016.
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4

Multiparameter Flow Cytometry of Immune Cells

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The following cell surface antibodies (purchased from eBioscience, San Diego, CA) and their respective isotype controls were used: anti-CD8α (53-6.7), anti-CD27 (LG.7F9), anti-CD45.1 (A20), anti-CD45.2 (104), anti-CD107a (1D4B), anti-Thy1.1 (HIS51) anti-PD-1 (J43) and anti-OX40 (OX86). Anti-Vβ8.3 TCR (1B3.3) was purchased from BD Biosciences (Oxford, UK). Direct intracellular staining was carried out using anti-perforin (OMAK-D), anti-T-bet (4B10) and anti-Eomes (Dan11mag) and isotype controls, purchased from eBioscience, San Diego, CA or anti-granzyme B (GRB05) from Life Technologies, UK. Following brief peptide re-stimulation, intracellular staining was performed using anti-IFN-γ (XMG1.2) or anti-TNF-α (MP6-XT22) together with the appropriate isotype controls (BD Biosciences, Oxford, UK). Intra-nuclear staining for BrdU was carried out using anti-BrdU-APC flow kit (BD Biosciences, Oxford, UK), according to the manufacturer's instructions. To detect CD107a cells were restimulated for 4 hours in the presence or absence of 1μM UTY peptide with Golgi-Stop (BD Pharmingen) in the presence of anti-CD107a or isotype control. Cells were then re-surface stained with anti-CD107a or isotype. Flow cytometric analysis was performed on a LSRFortessa or FACs Canto II (BD Biosciences) and cell counting performed on a Coulter Counter (Beckman Coulter).
Buchan S., Manzo T., Flutter B., Rogel A., Edwards N., Zhang L., Sivakumaran S., Ghorashian S., Carpenter B., Bennett C., Freeman G.J., Sykes M., Croft M., Al-Shamkhani A, & Chakraverty R. (2014). OX40- and CD27-mediated co-stimulation synergize with anti-PD-L1 blockade by forcing exhausted CD8+ T cells to exit quiescence. Journal of immunology (Baltimore, Md. : 1950), 194(1), 125-133.
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5

Comprehensive Immunophenotyping by Flow Cytometry

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The following fluorochrome-conjugated antibodies were used for flow cytometry. Anti-CD3ε (Clone ID, 145-2C11), anti-CD5 (53-7.3), anti-CD19 (1D3), anti-B220 (RA3-6B2), anti-CD11b (M1/70), anti-CD 11c (N418), anti-NK1.1 (PK136), anti-TCRγδ (eBioGL3), anti-Gr-1 (RB6-8C5), anti-FcεR1 (MAR-1), anti-CD4 (RM4-5), anti-CD8a (53-6.7), anti-CD49b (DX5), anti-TER119 (TER-119), anti-IL-7Rα (A7R34), anti-Thy1.2 (30-H12), anti-CD44 (IM7), anti-c-Kit (2B8), anti-Sca-1 (D7), anti-CD45.1 (A20), anti-CD45.2 (104), anti-IL-13 (eBio13A), anti-IFN-γ (XMG1.2), anti-IL-5 (TRFK5), anti-IL-17A (eBio17B7), anti-RORγt (AFKJS-9) and anti-T-bet (4B10) antibodies were from eBioscience. Anti-KLRG1 (2F1), anti-IL-4 (11B11), anti-Ki67 (B56) and anti-GATA-3 (L50-823) antibodies were from BD Biosciences. Anti-ST2 antibody (DJ8) was from MD Bioproducts. Anti-IL-17RB antibody (752101) was from R&D Systems. recombinant IL-25 and IL-33 were from R&D Systems. For in vitro cell culture, recombinant IL-2, IL-7, IL-3, SCF, IL-4, IL-12, IL-1β, IL-6, IL-23 and TGF-β were from Peprotech or R&D Systems, and neutralizing antibodies, including anti-IL-4 (11B11), anti-IL-12 (C17.8) and anti-IFN-γ (XMG1.2), were from Harlan Laboratories. LIVE/DEAD fixable dead cell stain kit was from Life Technologies.
Huang Y., Guo L., Qiu J., Chen X., Hu-Li J., Siebenlist U., Williamson P.R., Urban JF J.r, & Paul W.E. (2014). IL-25-responsive, lineage-negative KLRG1hi cells are multipotential “inflammatory” type 2 innate lymphoid cells. Nature immunology, 16(2), 161-169.
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6

Multiparameter Immune Cell Phenotyping

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Single-cell suspensions were generated from spleen and lymph nodes as designated. Cells were stained with the following extracellular antibodies: anti-CD4 (RM4-5), anti-CD8 (53-6.7), anti-CD44 (IM7), anti-CXCR3 (CXCR3-173), anti-CD3ε (145-2C11), anti-CD45.1 (A20), anti-CD45.2 (104), anti-KLRG1 (2F1), and anti-CD127 (A7R34). Antibodies were purchased from eBioscience, BD, or Tonbo. LCMV NP118–126 and L. monocytogenes LLO91–99 tetramers were provided by the National Institutes of Health Tetramer Facility. For intracellular staining, cells were fixed with the FoxP3 Fix/Perm kit (eBioscience) and stained with the following antibodies: anti-Eomes (Dan11mag), anti–T-bet (4B10), anti–IFN-γ (XMG1.2), and anti-TNF (MP6-XT22).
Renkema K.R., Lee J.Y., Lee Y.J., Hamilton S.E., Hogquist K.A, & Jameson S.C. (2016). IL-4 sensitivity shapes the peripheral CD8+ T cell pool and response to infection. The Journal of Experimental Medicine, 213(7), 1319-1329.
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7

Immunological Profiling with Flow Cytometry

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The following antibodies for flow cytometry were from BD Biosciences: anti-B220 (RA3-6B2), anti-CD3 (2C11), anti-CD4 (RM4–5), anti-CD8 (Ly-3), anti-CD25 (PC61), anti-CD44 (IM7), anti-CD124 (mIL4R-M1), anti-IFN-γ (XMG1.2), anti-IL-17a (TC11-18H10), and anti-human CD8 (RPA-T8). Anti-CD62L (MEL-14), anti-IL-13 (eBio13A), anti-IL-5 (TRFK5), anti-T-bet (eBio4B10) and anti-Foxp3 (FJK-16s) were from eBioscience.
The following antibodies for immunoblotting were from Cell Signaling Technologies: Antibody to Rictor (2140), NDRG1 phosphorylated at Thr346 (5482), total NDRG1 (9408), antibody to Akt phosphorylated at S473 (4060), pan-Akt (4685), NEDD4-2 phosphorylated at Ser342 (4080), total NEDD4-2 (4013), TCF-1 (2206), antibody to β-catenin (9562), antibody to phosphorylated β-catenin (9561), antibody to phosphorylated STAT6 (9361). Anti-T-bet (4B10) and anti-GATA-3 (TWAJ) were from eBioscience. Anti-JunB (210) was from Santa Cruz Biotechnology. Anti-ubiquitin was and actin were from Sigma. MG132 was from Sigma, PP242 was from Calbiochem, and CFSE was from Invitrogen.
Heikamp E.B., Patel C.H., Collins S., Waickman A., Oh M.H., Sun I.H., Illei P., Sharma A., Naray-Fejes-Toth A., Fejes-Toth G., Sen J., Horton M.R, & Powell J.D. (2014). The AGC kinase serum- and glucocorticoid-regulated kinase 1 (SGK1) regulates TH1 and TH2 differentiation downstream of mTORC2. Nature immunology, 15(5), 457-464.
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8

Comprehensive Immunophenotyping by Flow Cytometry

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The following fluorochrome-conjugated antibodies were used for flow cytometry. Anti-CD3ε (Clone ID, 145-2C11), anti-CD5 (53-7.3), anti-CD19 (1D3), anti-B220 (RA3-6B2), anti-CD11b (M1/70), anti-CD 11c (N418), anti-NK1.1 (PK136), anti-TCRγδ (eBioGL3), anti-Gr-1 (RB6-8C5), anti-FcεR1 (MAR-1), anti-CD4 (RM4-5), anti-CD8a (53-6.7), anti-CD49b (DX5), anti-TER119 (TER-119), anti-IL-7Rα (A7R34), anti-Thy1.2 (30-H12), anti-CD44 (IM7), anti-c-Kit (2B8), anti-Sca-1 (D7), anti-CD45.1 (A20), anti-CD45.2 (104), anti-IL-13 (eBio13A), anti-IFN-γ (XMG1.2), anti-IL-5 (TRFK5), anti-IL-17A (eBio17B7), anti-RORγt (AFKJS-9) and anti-T-bet (4B10) antibodies were from eBioscience. Anti-KLRG1 (2F1), anti-IL-4 (11B11), anti-Ki67 (B56) and anti-GATA-3 (L50-823) antibodies were from BD Biosciences. Anti-ST2 antibody (DJ8) was from MD Bioproducts. Anti-IL-17RB antibody (752101) was from R&D Systems. recombinant IL-25 and IL-33 were from R&D Systems. For in vitro cell culture, recombinant IL-2, IL-7, IL-3, SCF, IL-4, IL-12, IL-1β, IL-6, IL-23 and TGF-β were from Peprotech or R&D Systems, and neutralizing antibodies, including anti-IL-4 (11B11), anti-IL-12 (C17.8) and anti-IFN-γ (XMG1.2), were from Harlan Laboratories. LIVE/DEAD fixable dead cell stain kit was from Life Technologies.
Huang Y., Guo L., Qiu J., Chen X., Hu-Li J., Siebenlist U., Williamson P.R., Urban JF J.r, & Paul W.E. (2014). IL-25-responsive, lineage-negative KLRG1hi cells are multipotential “inflammatory” type 2 innate lymphoid cells. Nature immunology, 16(2), 161-169.
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