MIP-2 cell line

MIP-2 Leeporter Luciferase Reporter-RAW264.7 Cell Line

Cat. Num. AAA669505

• Applications: Functional Assay
• Synonyms: MIP-2; MIP-2 Leeporter Luciferase Reporter-RAW264.7 Cell Line; MIP-2 cell line

• Host: RAW264.7 cells
• Form/Format: Each vial contains 2-3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.

• Applicable Applications for MIP-2 cell line: Functional Assay (FA)
• Application Notes: Monitor the MIP-2 induction activity. Screen for activators or inhibitors of the MIP-2 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 1~2 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. 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 MIP-2 Leeporter - RAW264.7 cells to lipopolysaccharide (LPS).; 1. Harvest MIP-2 Leeporter - RAW264.7 cells and seed cells into a white solid-bottom 96-well microplate in 100 ul of growth medium at 8.5 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 different 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.

Related Product Information for MIP-2 cell line

The MIP-2 Leeporter Luciferase Reporter cell line is a stably transfected RAW264.7 cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the MIP-2 promoter. Macrophage inflammatory protein 2 (MIP-2) is a small cytokine that belongs to the C-X-C chemokine family and is also known as CXCL2. MIP-2 is one of the major proinflammatory cytokines, which is induced by innate immune receptors such TLRs and Nods, and also mediates LPS-induced osteoclastogenesis. The MIP-2 induction by Toll-like receptor 4 (TLR4) ligand, LPS, is shown in Figure 1.

Product Categories/Family for MIP-2 cell line

Reporter Cell Lines