IL-6 Reporter – NIH3T3 Cell Line

Product code: 14-116ACL

Application : Functional Assay

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2000 (throughout India)
Order now and get it on next 3-4 working days* (Major Cities)

Size
Price
1 Vial
70,000.00 


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Shipping Info:

2000 (throughout India)

Order now and get it on next 3-4 working days* (Major Cities)


Amount : 1 Vial

The IL-6 reporter cell line is a stably transfected NIH3T3 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 TLR ligands. The IL-6 induction by LPS 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 IL-6 induction activity.
  • Screen for activators or inhibitors of the IL-6 signaling pathway. 


Culture conditions:


Cells should be grown at 37oC with 5% CO2 using DMEM medium supplemented with 10% FBS and 1% Pen/Strep, plus 3 µg/ml of Puromycin.

It is recommended to quickly thaw the frozen cells upon receipt or from liquid nitrogen in a 37oC 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 37oC-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.
 

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 µl of growth medium at 5 x 104 cells/well. 

2. Incubate cells at 37oC in a CO2 incubator for overnight.
 
3. The next day, stimulate cells with various concentrations of LPS. 
 
4. Incubate at 37oC in a COincubator for 6-16 hours.
 
5. Add 50 µl of  luciferase assay reagent (Abeomics, Cat #17-1101) per well. 
 
6. Incubate at room temperature for 1-5 minutes and 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 is solely for Internal Research Purposes and not 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.

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

 

 


IL-6 Reporter – NIH3T3 Cell Line

Product code: 14-116ACL
mg

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