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Product Info Summary
| SKU: | RC1000 |
|---|---|
| Size: | 1 Vial, Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO. |
| Application: | Functional Assay |
Product info
Product Name
NF-kB Luciferase Reporter-RAW264.7 Cell Line
SKU/Catalog Number
RC1000
Size
1 Vial, Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.
Description
The NF-kB Luciferase Reporter cell line is a stably transfected RAW 264.7 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the NF-kB response element. NF-kB is a key transcription factor that is involved in immune and inflammatory responses, developmental processes, cellular growth and apoptosis.
Contents
Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.
Storage & Handling
Immediately upon receipt, store in liquid nitrogen. (Ship on dry ice.)
Cite This Product
NF-kB Luciferase Reporter-RAW264.7 Cell Line (Boster Biological Technology, Pleasanton CA, USA, Catalog # RC1000)
Assay dilution & Images
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. Actual working concentration varies and should be decided by the user.
| Application | Dilution | Species |
|---|---|---|
| Screen for activators or inhibitors of the NF-kB signaling pathway. Culture conditions: Cells should be grown at 37°C with 5% CO2 using DMEM medium supplemented with 10% FBS and 1% Pen/Strep | plus 3 µg/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 1~2 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. Note: RAW264.7 cells may not be detached well by trypsinization only. So you may need to use a cell scraper to harvest the trypsinized cells. |
| 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 NF-kB RAW264.7 cells to lipopolysaccharide (LPS) |
| 3. The next day | stimulate cells with different concentrations of LPS. |
Validation Images & Assay Conditions
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Fig-1: Induction of NF-kB activity by LPS in NF-kB RAW264.7 cells.
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Fig-2: Induction of NF-kB activity by LPS in NF-kB Leeporter™ - RAW264.7 cells.
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Fig-3: Induction of NF-kB activity by various TLR ligands in NF-kB Leeporter™ - RAW264.7 cells. Pam3CSK4 (10 ng/ml), Poly(I).Poly(C)-HMW (50 ug/ml), LPS (100 ng/ml), Flagellin (100 ng/ml), R848 (10 ug/ml), and CpG ODN 1826 (10 ug/ml).
Specific Publications For NF-kB Luciferase Reporter-RAW264.7 Cell Line (RC1000)
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