HRE Reporter – HeLa Cell Line

Product code: 14-118ACL

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 HRE reporter cell line is a stably transfected HeLa cell line which expresses Renilla luciferase reporter gene under the transcriptional control of the hypoxia response element (HRE). In response to hypoxia (low oxygen), HREs of target genes are recognized and regulated by the hypoxia-inducible factors (HIFs) which belong to the family of basic helix-loop-helix transcription factors and form heterodimeric complex comprising the alpha subunit (HIF-1 alpha, HIF-2 alpha and HIF-3 alpha) and beta subunit (Arnt1, Arnt2 and Arnt3), among which HIF-1 alpha and HIF-2 alpha are predominant isoforms. Activation of HIFs can also be mediated by chemical hydroxylase inhibitors as hypoxia mimetics including the iron chelator desferrioxamine and cobalt chloride.

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 HIF induction activity.
  • Screen for activators or inhibitors of the hypoxia signaling pathway. 

Culture conditions:

Cells should be grown at 37⁰C 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 2~4 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.

Functional validation:

A. Response of HRE Leeporter™ – HeLa cells to cobalt chloride (CoCl2).
 
1. Harvest HRE Leeporter™ – HeLa cells and seed cells into a white solid-bottom 96-well microplate in 100 µl of growth medium at 5 x 10^4 cells/well. 
 
2. Incubate cells at 37oC in a CO2 incubator for overnight.
 
3. The next day, stimulate cells with various concentrations of CoCl2
 
4. Incubate at 37oC in a CO2 incubator 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).

 


HRE Reporter – HeLa Cell Line

Product code: 14-118ACL
mg

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