Application Data
(Fig-1: . Induction of GATA3 activity by PMA in GATA3 Leeporter HEK293 cells.)
GATA3 cell line
GATA3 Leeporter HEK293 Cell Line
Gene Names
GATA3; cGATA-3
Applications
Functional Assay
Synonyms
GATA3; N/A; GATA3 Leeporter HEK293 Cell Line; GATA3 cell line
Form/Format
Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.
Sequence Length
444
Applicable Applications for GATA3 cell line
FA (Functional Assay)
Culture conditions
Cells should be grown at 37°C with 5% CO2 using DMEM medium (w/ L-Glutamine, 4.5g/L Glucose and Sodium Pyruvate) supplemented with 10% heat-inactivated FBS and 1% Pen/Strep, plus 3 ug/ml of Puromycin (Note: Puromycin can be omitted during the reporter cell assays).
It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37°C water-bath, transfer to a tube containing 10 ml of growth medium without Puromycin, spin down cells, resuspend cells in pre-warmed growth medium without Puromycin, transfer resuspended cells to T25 flask and culture in 37°C-CO2 incubator.
Leave the T25 flask in the incubator for 1~3 days without disturbing or changing the medium until cells completely recover viability and become adherent. Once cells are over 90% adherent, remove growth medium and passage the cells through trypsinization and centrifugation. At first passage, switch to growth medium containing Puromycin. Cells should be split before they reach complete confluence.
To passage the cells, detach cells from culture vessel with Trypsin/EDTA, add complete growth medium and transfer to a tube, spin down cells, resuspend cells and seed appropriate aliquots of cells suspension into new culture vessels. Subcultivation ration = 1:10 to 1:20 weekly. To achieve satisfactory results, cells should not be passaged over 16 times.
It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37°C water-bath, transfer to a tube containing 10 ml of growth medium without Puromycin, spin down cells, resuspend cells in pre-warmed growth medium without Puromycin, transfer resuspended cells to T25 flask and culture in 37°C-CO2 incubator.
Leave the T25 flask in the incubator for 1~3 days without disturbing or changing the medium until cells completely recover viability and become adherent. Once cells are over 90% adherent, remove growth medium and passage the cells through trypsinization and centrifugation. At first passage, switch to growth medium containing Puromycin. Cells should be split before they reach complete confluence.
To passage the cells, detach cells from culture vessel with Trypsin/EDTA, add complete growth medium and transfer to a tube, spin down cells, resuspend cells and seed appropriate aliquots of cells suspension into new culture vessels. Subcultivation ration = 1:10 to 1:20 weekly. To achieve satisfactory results, cells should not be passaged over 16 times.
Functional validation
A. Response of GATA3 Leeporter™ – HEK293 cells to phorbol 12-myristate 13-acetate (PMA).
1. Harvest GATA3 Leeporter™ – HEK293 cells and seed cells into a white solid-bottom 96-well microplate in 100 ul of growth medium at 5 x 10^4 cells/well.
2. Incubate cells at 37°C in a CO2 incubator for overnight.
3. The next day, stimulate cells with different concentrations of PMA.
4. Incubate at 37°C in a CO2 incubator for 16 hours.
5. Equilibrate the plate to room temperature for 10 minutes.
6. Add 50 ul of luciferase assay reagent (MBS668917; Refer to the reagent datasheet for the detailed luciferase assay protocol) per well.
7. Read the plate in 1-5 minutes to measure luminescence using a microplate luminometer.
1. Harvest GATA3 Leeporter™ – HEK293 cells and seed cells into a white solid-bottom 96-well microplate in 100 ul of growth medium at 5 x 10^4 cells/well.
2. Incubate cells at 37°C in a CO2 incubator for overnight.
3. The next day, stimulate cells with different concentrations of PMA.
4. Incubate at 37°C in a CO2 incubator for 16 hours.
5. Equilibrate the plate to room temperature for 10 minutes.
6. Add 50 ul of luciferase assay reagent (MBS668917; Refer to the reagent datasheet for the detailed luciferase assay protocol) per well.
7. Read the plate in 1-5 minutes to measure luminescence using a microplate luminometer.
Dry Ice Shipment
Extra charge fee may add to your shipping cost as dry ice is required to ship this product.
Preparation and Storage
Immediately upon receipt, store in liquid nitrogen.
Dry Ice Shipment: Extra charge fee may add to your shipping cost as dry ice is required to ship this product.
Shipping Note: Product is available for shipment in the United States, Canada and European countries. Please inquire for shipment to other countries.
Dry Ice Shipment: Extra charge fee may add to your shipping cost as dry ice is required to ship this product.
Shipping Note: Product is available for shipment in the United States, Canada and European countries. Please inquire for shipment to other countries.
Application Data
(Fig-1: . Induction of GATA3 activity by PMA in GATA3 Leeporter HEK293 cells.)
Application Data
(Fig-1: . Induction of GATA3 activity by PMA in GATA3 Leeporter HEK293 cells.)
Related Product Information for GATA3 cell line
The GATA3 Leeporter™ Luciferase Reporter cell line is a stably transfected HEK 293 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the GATA3 response element, so that the cell line is designed to measure the transcriptional activity of GATA3. As a zinc-finger transcription factor, GATA3 (GATA-binding protein 3) plays a critical role in early and late T cell differentiation, which regulates Th1/Th2 differentiation. GATA3 has been shown to induce Th2 differentiation and repress Th1 differentiation. GATA3 is also known to promote the secretion of IL-4, IL-5 and IL-13 from Th2 cells. The GATA3 induction by phorbol 12-myristate 13-acetate (PMA) is shown in Figure 1.
Product Categories/Family for GATA3 cell line
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
NCBI Official Full Name
GATA-binding factor 3
NCBI Official Synonym Full Names
GATA binding protein 3
NCBI Official Symbol
GATA3
NCBI Official Synonym Symbols
cGATA-3
NCBI Protein Information
GATA-binding factor 3
UniProt Protein Name
GATA-binding factor 3
UniProt Gene Name
GATA3
UniProt Synonym Gene Names
GATA-3
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Product Notes
The GATA3 gata3 (Catalog #AAA78373) is a Cell Line and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's GATA3 can be used in a range of immunoassay formats including, but not limited to, FA (Functional Assay). Researchers should empirically determine the suitability of the GATA3 gata3 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "GATA3, Cell Line" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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