BM, splenocytes, and LN cells were stained with fluorescent conjugated Abs specific for CD4 (GK1.5, Tonbo Biosciences), CXCR5 (L138D7, BioLegend), PD1 (29F.1A12, BioLegend), CD38 (90, BioLegend), GL7 (GL7, BioLegend), B220 (RA3-6B2, BioLegend), CD93 (AA4.1, BioLegend), CD19 (1D3, Tonbo Biosciences), CD21 (7E9, BioLegend), CD23 (BEB4, BioLegend), CD24 (M1/69, BioLegend), CD62L (MEL-14, Tonbo Biosciences), CD25 (7D4, Tonbo Biosciences), CD43 (S7, BD Pharmigen), IgD (11-26C.2A, BioLegend), and IgM (DS-1, BD Pharmigen); Igκ (1050-02; Southern Biotech); T-bet (EBIO4B10, eBiosciences) and Bcl6 (K112-91, eBioscience); and BrdU (BU20A, BioLegend). Other reagents include Caspase3/7 (C10427, Invitrogen), ghost dye (13-0865-T100, Tonbo Biosciences), and NP-PE (N-5070-1; Biosearch Technologies). Cells were analyzed by flow cytometry as previously described (86 (link)). VLP Ab was a gift from Shaun Jackson. Intracellular Bcl6 staining to identify Tfh cells was performed as previously described (87 (link)). Intracellular staining for T-bet was performed as previously described (23 (link)).
T bet ebio4b10
Manufactured by Thermo Fisher Scientific
The T-bet (EBIO4B10) is a laboratory equipment product from Thermo Fisher Scientific. It is a transcription factor that plays a critical role in the differentiation and function of T helper 1 (Th1) cells, which are involved in cell-mediated immune responses. The T-bet (EBIO4B10) is a tool that can be used in research and scientific applications to study the regulatory mechanisms of T cell development and function.
Automatically generated - may contain errors
Lab products found in correlation
Facscanto 2, by BD (2 mentions)
Cd38 90, by BioLegend (1 mentions)
B220 ra3 6b2, by BioLegend (1 mentions)
Cd93 aa4, by BioLegend (1 mentions)
Cd19 1d3, by Cytek Biosciences (1 mentions)
Cd21 7e9, by BioLegend (1 mentions)
Cd24 m1 69, by BioLegend (1 mentions)
Igd 11 26c 2a, by BioLegend (1 mentions)
Pd 1 29f 1a12, by BioLegend (1 mentions)
7 amino actinomycin d 7 aad viability staining solution, by BioLegend (1 mentions)
C maf sym0f1, by Thermo Fisher Scientific (1 mentions)
Facsaria 2, by BD (1 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
Facsaria instrument, by BD (1 mentions)
Anti pd 1 rmp1 30, by BioLegend (1 mentions)
Eomes dan11mag, by Thermo Fisher Scientific (1 mentions)
Klrg1 2f1, by Thermo Fisher Scientific (1 mentions)
Granzyme b gb12, by Thermo Fisher Scientific (1 mentions)
Transcription factor staining kit, by Thermo Fisher Scientific (1 mentions)
Cytofix cytoperm kit, by BD (1 mentions)
6 protocols using t bet ebio4b10
1
Multiparametric Flow Cytometry of Immune Cells
2
Profiling Peripheral Blood Mononuclear Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fresh peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation using Ficoll-Paque™ PLUS (GE Healthcare Bioscience) and stained for cell surface antigens employing the following fluorescently labelled monoclonal and isotype control antibodies: CD45RA (HI100), CD3 (UCHT1, OKT3), CD4 (OKT4), CD8α (HIT8a, RDA-T8), CCR9 (L053E8), CCR6 (G034E3), CCR7 (G043H7), β1 (TS2/16), β7 (FIB504) (BioLegend), α4 (9F10) (BD), CXCR3 (49801) (R&D). Dead cells were stained with the 7-aminoactinomycin D (7-AAD) viability staining solution (BioLegend). In some experiments, intracellular staining was performed with antibodies against RORγt (AFKJS-9), T-BET (eBio4B10), and C-MAF (sym0F1) (eBioscience) using the Cytofix or Cytoperm™ Kit (Becton Dickinson). Mouse or rat serum (eBioscience) was used for blocking. Intracellular FOXP3 was stained using the PE anti-human FOXP3 staining kit (eBioscience). Cells were analysed or sorted by BD FACSCanto II or FACSAria II.
3
Comprehensive Immune Profile of T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Surface staining was performed for 30 min and with tetramer for 90 min at 4°C in PBS supplemented with 2% FCS and 0.01% azide (FACS buffer) using the following antibodies: anti-CD8α (53-6.7), CD127 (eBioSB/199), LAG-3 (C9B7W), and KLRG-1 (2F1; all eBioscience); anti–PD-1 (RMP1-30; BioLegend); gp276–286 tetramer (TCMetrix); and anti-CD4 (GK1.5), CD45.1 (A20), and CD45.2 (clone 104), obtained from or custom purified by Bio X Cell and coupled to Pacific blue, Alexa Fluor 647, or FITC using labeling reagents from Invitrogen. Cells were washed twice and fixed in PBS supplemented with 1% formaldehyde, 2% glucose, and 0.03% azide for 20 min. Then, cells were washed again and resuspended in FACS buffer. For intracellular cytokine staining, cells were fixed and permeabilized using the Cytofix/Cytoperm kit (BD) and stained with anti–IFN-γ (XMG1.2), TNF (MP6-XT22; both from eBioscience), or granzyme B (GB12; Invitrogen). T-bet and Eomes staining was performed with a transcription factor staining kit (eBioscience) and stained with Eomes (Dan11mag) and T-bet (eBio4B10; both from eBioscience).
For flow cytometry sorting, living cells were stained in 10% FCS RPMI media and sorted on a FACSAria instrument (BD).
For flow cytometry sorting, living cells were stained in 10% FCS RPMI media and sorted on a FACSAria instrument (BD).
4
Multiparametric Flow Cytometry of Thymocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Surface staining was performed for 20 minutes at room temperature and always included Fixable Viability Dye (eBioscience) for dead cell exclusion. For intracellular analysis thymocytes were fixed, permeabilized and stained using the Transcription Factor Staining Buffer Set (eBioscience), as per manufacturer’s instructions. Other anti-human mAbs used included: CD8β (SIDI8BEE), CD27 (O323), CD45RA (HI100), CD44 (IM7), CD69 (FN50), CXCR3 (1C6), Eomes (WD1928), T-bet (eBio4B10), ThPOK (ZFP-67), Runx1 (RXDMC), Runx3 (R3-5G4) and Ki67 (B56) from eBioscience/Thermo Fisher Scientific, BD Biosciences or R&D Systems. Cells were acquired in a LSRFortessa (BD Biosciences) cytometer and data was analyzed using FlowJo v10 (FlowJo, BD). For the visualization of high-dimensional data through the t-distributed stochastic neighbor embedding (t-SNE) dimensionality reduction technique, similar number of live, single cells from each condition were concatenated. tSNE parameters were set to 1,000 iterations, perplexity 30 and learning rate 4200, and based on appropriate markers.
5
Comprehensive Immune Cell Phenotyping
6
Multiparameter Flow Cytometry Immunophenotyping
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Single-cell suspensions in PBS 1 × were stained with fixable viability dye eFluro™ 620 (eBioscience). Cells were preincubated with anti-CD16/32 for 10 min to block Fc receptors and stained in FACS buffer (PBS containing 1% BSA) with directly labeled monoclonal antibodies for 30 min. For intracellular cytokine staining, cells were activated for 4 h with 50 ng/ml PMA, 1 μg/ml ionomycin in the presence of 10 mg/ml brefeldin A. After surface staining, cells were fixed and permeabilized using Foxp3/transcription factor staining buffer set and stained intracellularly with directly labeled monoclonal antibodies for 30 min. Data were acquired on LSR II cytometer and all data were analyzed using FlowJo software. Fluorochrome-conjugated antibodies were purchased from several commercial sources indicated below. Antibodies against CD45 (30-F11) was from Biolegend; CD3 (145-2C11), CD4 (GK1.5), IL-17A (Q31-378), RORγt (ebio17B7), T-bet (ebio4B10) and IFN-γ (XMG1.2) were from eBiosciences.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!