Testing Data
(Fig-1: Induction of CRE activity by forskolin in CRE Leeporter HEK293 cells.)
CRE cell line
CRE Leeporter HEK293 Cell Line
Applications
Functional Assay
Synonyms
CRE; CRE Leeporter HEK293 Cell Line; CRE cell line
Form/Format
Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.
Applicable Applications for CRE cell line
Functional Assay
Application Notes
Monitor the GPCR-linked cAMP/PKA signaling pathway. Screen for activators or inhibitors of the GPCR-linked cAMP/PKA signaling pathway. Culture conditions: Cells should be grown at 37 degree C with 5% CO2 using DMEM medium supplemented with 10% FBS and 1% Pen/Strep, plus 3 ug/ml of Puromycin. It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37 degree 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 degree C-CO2 incubator. Leave the T25 flask in the incubator for 2~4 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. Functional validation: A. Response of CRE Leeporter- HEK293 cells to forskolin. 1. Harvest CRE 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 degree C in a CO2 incubator for overnight. 3. The next day, stimulate cells with various concentrations of the forskolin. 4. Incubate at 37 degree C in a CO2 incubator for 6-16 hours. 5. Add 30-50 ul of luciferase assay reagent per well. 6. Incubate at room temperature for 1-5 minutes and measure luminescence using a microplate luminometer.
Shipping Note
Product available only in the USA.
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.
Testing Data
(Fig-1: Induction of CRE activity by forskolin in CRE Leeporter HEK293 cells.)
Testing Data
(Fig-1: Induction of CRE activity by forskolin in CRE Leeporter HEK293 cells.)
Related Product Information for CRE cell line
The CRE Leeporter cell line is a stably transfected HEK 293 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the cAMP response element (CRE). The CRE Leeporter cell line is designed to monitor the cAMP/PKA signaling pathways and can be used for studying GPCR-linked cAMP/PKA signaling pathways as well as screening of agonists, antagonists or signaling inhibitors related with the cAMP/PKA signaling pathways. Functional activity of the cell line has been validated by serum stimulation assay.
Product Categories/Family for CRE cell line
Similar Products
Product Notes
The CRE (Catalog #AAA668850) is a Cell Line and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's CRE can be used in a range of immunoassay formats including, but not limited to, Functional Assay. Monitor the GPCR-linked cAMP/PKA signaling pathway. Screen for activators or inhibitors of the GPCR-linked cAMP/PKA signaling pathway. Culture conditions: Cells should be grown at 37 degree C with 5% CO2 using DMEM medium supplemented with 10% FBS and 1% Pen/Strep, plus 3 ug/ml of Puromycin. It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37 degree 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 degree C-CO2 incubator. Leave the T25 flask in the incubator for 2~4 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. Functional validation: A. Response of CRE Leeporter- HEK293 cells to forskolin. 1. Harvest CRE 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 degree C in a CO2 incubator for overnight. 3. The next day, stimulate cells with various concentrations of the forskolin. 4. Incubate at 37 degree C in a CO2 incubator for 6-16 hours. 5. Add 30-50 ul of luciferase assay reagent per well. 6. Incubate at room temperature for 1-5 minutes and measure luminescence using a microplate luminometer. Researchers should empirically determine the suitability of the CRE 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 "CRE, 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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