IL-6 cell line

IL-6 Leeporter Luciferase Reporter-NIH 3T3 Cell Line

Cat. Num. AAA669517

• Applications: Functional Assay
• Synonyms: IL-6; IL-6 Leeporter Luciferase Reporter-NIH 3T3 Cell Line; IL-6 cell line

• Host: NIH 3T3 cells
• Form/Format: Each vial contains 2-3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.

• Applicable Applications for IL-6 cell line: Functional Assay (FA)
• Application Notes: Monitor the IL-6 induction activity. Screen for activators or inhibitors of the IL-6 signaling pathway.
• Culture Conditions: Cells should be grown at 37 degree 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 3ug/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 degree C water-bath, transfer to a tube containing 10ml 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 1~3 days without disturbing or changing the medium until cells completely recover viability and become adherent. Once cells are between 80~90% adherent, remove growth medium and passage the cells through trypsinization and centrifugation. At first passage, switch to growth medium containing Puromycin.; Note: NIH 3T3 cells should be split before they reach 90% confluence; otherwise, they become self-lifted and aggregate irrevisibly. Precoating the cell assay plates with 0.1% gelatin may prevent NIH 3T3 cells from self-lifting. 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 IL-6 Leeporter - NIH 3T3 cells to lipopolysaccharide (LPS).; 1. Harvest IL-6 Leeporter - NIH 3T3 cells and seed cells into a white solid-bottom 96-well microplate in 100 ul of growth medium at 5 x 104 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 LPS.; 4. Incubate at 37 degree C in a CO2 incubator for 6-16 hours.; 5. Add 50 ul of luciferase assay reagent (Cat #MBS668917) 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.

Activity

(Fig-1: Induction of IL-6 promoter activity by LPS in IL-6 Leeporter™ – NIH 3T3 cells.)

Activity

(Fig-1: Induction of IL-6 promoter activity by LPS in IL-6 Leeporter™ – NIH 3T3 cells.)

Related Product Information for IL-6 cell line

The IL-6 Leeporter Luciferase Reporter cell line is a stably transfected NIH 3T3 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the IL-6 promoter. As a pleiotropic cytokine, interleukin 6 (IL-6) has pro- and anti-inflammatory roles which is not only involved in normal functions of the immune system, hematopoiesis and  metabolism but also involved in the pathogenesis of metabolic and cardiovascular diseases. IL-6 gene induction is generally regulated by several transcription factors that activate the consensus sequences in the IL-6 promoter region, which include AP-1, C/EBP-beta and NF-kB in response to various proinflammatory cytokines, growth factors, and pathogen-associated molecular patterns such as Toll-like receptor (TLR) ligands. The IL-6 induction by lipopolysaccharide (LPS), the TLR4 ligand, is shown in Figure 1.

Product Categories/Family for IL-6 cell line

Reporter Cell Lines