安倍经济学/肿瘤坏死因子-α™ 荧光素酶报告基因-RAW264.7细胞系/营利性/14-106ACL-免疫在线蚂蚁淘旗下平台

商品详细安倍经济学/肿瘤坏死因子-α™ 荧光素酶报告基因-RAW264.7细胞系/营利性/14-106ACL

安倍经济学/肿瘤坏死因子-α™ 荧光素酶报告基因-RAW264.7细胞系/营利性/14-106ACL

商品编号： 14-106ACL

品牌： Abeomics

市场价： ¥0.00

美元价： 0.00

产地：美国(厂家直采)

公司：

产品分类： WB试剂盒

公司分类： Westornblot

联系Q Q： 3392242852

电话号码： 4000-520-616

电子邮箱： info@ebiomall.com

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Amount :	Non Profit

The TNF-alpha 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 TNF-alpha promoter. Tumor necrosis factor-alpha (TNF-alpha) is one of the major proinflammatory cytokines and can induce systemic inflammation, apoptotic cell death, sepsis and cachexia. Dysregulation of TNF-alpha induction is often involved in various human diseases including inflammatory bowel disease, cancer and Alzheimer's disease. The TNF-alpha induction by lipopolysaccharide (LPS), the Toll-like receptor 4 (TLR4) ligand, is shown in Figure 1.

Content :	Each vial contains 2 ~ 3 x 10^6 cells in 1 ml of 90% FBS + 10% DMSO.
Storage condition :	Immediately upon receipt, store in liquid nitrogen.

Application:

Monitor the TNF-alpha induction activity.
Screen for activators or inhibitors of the TNF-alpha signaling pathway.

Culture conditions:

Cells should be grown at 37^oC with 5%CO₂using DMEM medium (w/ L-Glutamine, 4.5g/L Glucose and Sodium Pyruvate) supplemented with 10% heat-inactivated FBS and 1% Pen/Strep, plus 3µg/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 a37^oCwater-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 in37^oC-CO₂incubator.

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.To achieve satisfactory results, cells should not be passaged over 16 times.

Functional validation:A. Response of TNF-alpha Leeporter™ – RAW264.7 cells to lipopolysaccharide (LPS).

1. Plate TNF-alpha Leeporter™ –RAW264.7 cells into a white solid-bottom 96-well microplate in 100µlof growth medium at 1 x 10⁵cells/well and incubate cells at37^oCin a CO₂incubator for 4-6 hours.

2. Stimulate cells with different concentrations of LPS and incubate cells at37^oCin a CO₂incubator for 16 hours.

3. Equilibrate the plate to room temperature for 10 minutes.

4. Add 50 µl of luciferase assay reagent (Abeomics, Cat #17-1101; Refer to the reagent datasheet for the detailed luciferase assay protocol) per well.

5. Read the plate in 1-5 minutes to measure luminescence using a microplate luminometer.

LIMITED USE RESTRICTIONS:

THIS PRODUCT IS SOLELY FOR IN VITRO RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE.

By use of this product, user agrees to be bound by the terms of this limited use statement.

This product issolely for Internal Research Purposesandnot for Commercial Purposes. Commercial Purposes include, but are not limited to (1) use of the cell line in manufacturing; (2) use of the cell line to provide a service, information or data; (3) use of the cell line for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the cell line whether or not such cell lines are resold for use in research.The buyer cannot sell, give or otherwise transfer this product to a third party.

Commercial License Agreement is available for non-research use if applicable. Please contact Abeomics (info@abeomics.com).

For Research Use Only. Not for use in diagnostic/therapeutics procedures.

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