Gata 3

Manufactured by BioLegend

Sourced in United States

GATA-3 is a transcription factor that plays a crucial role in the development and differentiation of T helper type 2 (Th2) cells. It is an essential regulator of Th2 cell lineage commitment and function.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Anti-human/mouse GATA-3 (TWAJ and 16E10A23)

6 protocols using gata 3

Multiparametric Flow Cytometry for Hematopoietic Stem Cell Analysis

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

The following monoclonal antibodies were used: Anti-Lineage, a cocktail containing a concentration optimized mix of the following antibodies: CD3e, Ly6G/Ly6c, CD11b, CD45R/B220, TER-119 (BioLegend, San Diego, CA). Monoclonal Antibodies against CD117 (c-kit), Sca-1, CD127 (IL7R), CD45, KLRG-1, CD25, CD90, CD34, ST2, IL17RB, CCR2, T-bet, Gata-3, and RORgt were purchased from BioLegend. Live/Dead Yellow Fixable Stain, Fixable Viability Dye eFluor 780 or DAPI were utilized to assess viability. Data acquisition and/or cell sorting was performed on a special-order 5-laser BD LSRII flow cytometer, Beckman Coulter Gallios, or BD FACS Aria or Aria II instrument. Purity after sorting was routinely >95%. The analysis was performed using FlowJo software. Mean fluorescence intensities (MFIs) were calculated using the geometric mean of the appropriate fluorescence channel in FlowJo. Expansion Indices were determined using the embedded FlowJo algorithm. For the magnetic separation of hematopoietic stem cells, we used the EasySep mouse CD117 (c-kit) positive selection kit from StemCell Technologies (18757) and we followed the protocol provided by the company.

Korniotis S., Thornley T.B., Kyriazis P., Theodorou E., Ma L., Li L.S., Kokkotou E., Strom T.B, & Koulmanda M. (2018). Hematopoietic Stem/Progenitor Cell Dependent Participation of Innate Lymphoid Cells in Low-Intensity Sterile Inflammation. Frontiers in Immunology, 9, 2007.

+ Open protocol

+ Expand

Comprehensive Immune Cell Profiling Protocol

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

Cells were incubated for 4 h with phorbol-12-myristate 13-acetate [21 (link)], ionomycin and brefeldin A, stained with a fixable viability stain (Zombie UV; BioLegend, London, UK) and ILC and T-cell markers. All ILCs were lineage-negative (CD3, CD14, CD16, CD19, CD20, CD56, CD4, FcεR1), CD45⁺. Type 2 ILCs were CRTH2⁺ or CD127⁺ and/or IL-13⁺ or IL-4⁺. Th2 cells were CD3⁺CD4⁺ expressing CRTH2/IL-13/IL-4. Th17 cells (CD4⁺IL-17⁺) and IL-17⁺ILCs (lineage-negative, IL-17⁺). Antibodies used were CD45 (Life Technologies, Paisley, UK), lineage cocktail, CD127, CRTH2, CD3, CD4, CD8, IL-13, IL-17A, GATA-3 (BioLegend) FcεR1 (eBioscience, Altringham, UK). Data was acquired on BD Fortessa (BD Bioscience, SanJose, CA, USA) and analysed using Flowjo v10 (Flowjo, Ashland, OR, USA).

Nagakumar P., Puttur F., Gregory L.G., Denney L., Fleming L., Bush A., Lloyd C.M, & Saglani S. (2019). Pulmonary type-2 innate lymphoid cells in paediatric severe asthma: phenotype and response to steroids. The European Respiratory Journal, 54(2), 1801809.

+ Open protocol

+ Expand

Comprehensive Immune Cell Analysis

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

Fluoroisothiocyanate, PE/dazzle, allophycocyanin, and phycoerythrin-labeled CD3, CD4, CD8, CXCR4, CXCR7, CD45R, HLA-DR, GATA3, Ki-67, Helios, and FOXP3 human monoclonal antibodies, RBC lysis buffer (10X), intracellular staining permeabilization wash buffer (10X), and fixation buffer were purchased from BioLegend (San Diego, USA). GolgiStop was purchased from BD Biosciences (San Diego, USA). RPMI 1640 medium, phorbol myristate acetate (PMA), and ionomycin were purchased from Sigma-Aldrich (St. Louis, MO, USA). TRIzol was purchased from Life Technologies (Paisley, UK). SYBR green and cDNA kits were purchased from Applied Biosystems (Foster City, CA, USA). Primers were synthesized from GenScript (Piscataway, NJ, USA).

Alhosaini K., Ansari M.A., Nadeem A., Attia S.M., Bakheet S.A., Al-Ayadhi L.Y., Mahmood H.M., Al-Mazroua H.A, & Ahmad S.F. (2021). Dysregulation of Ki-67 Expression in T Cells of Children with Autism Spectrum Disorder. Children, 8(2), 116.

+ Open protocol

+ Expand

Multiparametric Flow Cytometry Analysis

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

Cell surface molecular expression and intracellular cytokine production were evaluated using flow cytometry. FITC-conjugated anti-human CD14, CD4, anti-mouse F4/80, CD4, TNF-α, IFN-γ, eFluor® 488-conjugated anti-human Foxp3, anti-mouse Foxp3, TNF-α, IFN-γ, STAT-1 Phospho, PE-conjugated anti-human Tim-3, CD132, IL-17A, IL-10, T-bet, GATA-3, anti-mouse CD80, Tim-3, TGF-β1, IL-10, IL-13, T-bet, PE/CY7-conjugated anti-human CD4, TNF-α, TGF-β1, anti-mouse IL-10, IL-12/23, TNF-α, T-bet, F4/80, PerCP/Cy5.5-conjugated anti-mouse T-bet, IL-17A, APC-conjugated anti-human Tim-3, IFN-γ, IL-13, ROR-γt, Foxp3, anti-mouse F4/80, TNF-α, Tim-3, IL-10, ROR-γt, GATA-3, STAT-6 Phospho, Pacific Blue-conjugated anti-mouse STAT-1 Phospho, Brilliant Violet 421-conjugated anti-human IL-4, anti-mouse CD206, TGF-β1, IL-4, TNF-α, STAT-1 Phospho, Brilliant Violet 510-conjugated anti-mouse CD4, CD86, TNF-α, IFN-γ, Brilliant Violet 605-conjugated anti-mouse CD4, IL-17A (Biolegend, U.S.A.), and PerCP-eFluor-710-conjugated anti-mouse GATA-3, (eBioscience, San Diego, CA, USA) antibodies were used. For intracellular staining, cells were fixed and permeabilized using the Fix/Perm kit (Biolegend, U.S.A.). Flow cytometry was performed on a Beckman-Coulter CyAn ADP cytometer (Beckman-Coulter, U.S.A.) and analyzed with FlowJo software (Tree Star, Ashland, U.S.A.).

Li M., Sun F., Xu Y., Chen L., Chen C., Cui L., Qian J., Li D., Wang S, & Du M. (2022). Tim-3+ decidual Mφs induced Th2 and Treg bias in decidual CD4+T cells and promoted pregnancy maintenance via CD132. Cell Death & Disease, 13(5), 454.

+ Open protocol

+ Expand

Western Blot Analysis of TNBC Cell Lysates

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

Lysates from TNBC cells were prepared using 20mM Tris, pH 8, 135 mM NaCl, 1% NP-40, 10% glycerol, 1mM PMSF, and complete protease inhibitor cocktail (Roche) lysis buffer. Protein lysates were resolved by 4%-15% gradient SDS-polyacrylamide gel electrophoresis, transferred to PVDF membrane, and probed with the indicated primary antibodies: EZH2 (Cell Signaling), H3K27me3 (Clone 07-449, Millipore), H3K4me3 (Abcam), GATA3 (Clone 16E10A23, Biolegend), ERa (Clone 1D5, Thermo), b-Actin (Sigma Aldrich), and a-Tubulin (Proteintech). Membranes were then incubated with a peroxidase-conjugated correspondent secondary antibody and detected using enhanced chemiluminescence.

Yomtoubian S., Lee S.B., Verma A., Izzo F., Markowitz G., Choi H., Cerchietti L., Vahdat L., Brown K.A., Andreopoulou E., Elemento O., Chang J., Inghirami G., Gao D., Ryu S, & Mittal V. (2020). Inhibition of EZH2 Catalytic Activity Selectively Targets a Metastatic Subpopulation in Triple-Negative Breast Cancer. Cell reports, 30(3).

+ Open protocol

+ Expand

Multiparameter T cell phenotyping

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

Cell surface markers were stained for 30 min in the dark at 4 °C. Intracellular cytokine staining was performed using the ebioscience Foxp3 staining kit (ThermoFisher Scientific, Waltham MA) per manufacturer’s protocol. The following mouse antibodies from BioLegend (San Diego CA) were used: CD4 (GK1.5); CD8 (53-6.7); Tbet (4B10); Gata3 (16E10A23); Foxp3 (MF-14); IFNγ (XMG1.2); IL-10 (JES5-16E3); IL17 (TC11-18H10.1); and TGFβ (TW7-16B4). RORγt (AFKJS-9) was ordered from eBioscience (ThermoFisher Scientific). To show T cell reactivity, splenocytes were isolated from tumor bearing mice and cultured with 4T1 cells at a ratio of 5:1 (splenocytes to tumor cells) in the presence anti-CD107A/CD107B (ThermoFisher Scientific) and Monensin for 4 h. After 4 h, cells were stained for surface and intracellular markers described above. Flow cytometry analysis of T cell markers on human PBMCs was performed using the following clones: CD8 (RPA-T8); CD4 (SK3); Tbet (4B10); Ki67 (Ki67) from BioLegend; RORγt and granzyme B (GB11) from ThermoFisher Scientific.

Draganov D., Han Z., Rana A., Bennett N., Irvine D.J, & Lee P.P. (2021). Ivermectin converts cold tumors hot and synergizes with immune checkpoint blockade for treatment of breast cancer. NPJ Breast Cancer, 7, 22.

+ 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!