Human IFN alpha kit
No-wash kit to quantify released Human IFN alpha
The HTRF Human ADAR1 Detection Kit is designed to monitor the expression level of cellular ADAR1.
ADAR1 is one of two ADAR gene-coded enzymes. It binds to double-stranded RNA (dsRNA) and deaminates its adenosine into inosine (hypoxanthine). ADAR proteins act post-transcriptionally to alter RNA nucleotide contents. The deamination they operate interferes with the usual A:U pairing and destabilizes the RNA.
The loss of regulation of ADAR1 & ADAR2 is involved in the development and progression of multiple cancers, such as glioblastomas, melanomas, or acute leukemias. On top of their role in oncogenesis, ADAR enzymes are suspected of contributing to the aggravation of some infectious diseases like HIV and measles, but also of mental disorders such as depression, epilepsy, and schizophrenia.
HepG2 cells were plated at different cell densities under 80µl in a 96-well plate in complete culture medium, and incubated overnight at 37°C, 5% CO2.
After culture, the cells were treated with siRNA by adding 20µl of a mix of Lipofectamine® RNAiMax/siRNA for ADAR1, then incubated for 24h and 48h at 37°C, 5% CO2. For untreated cells, 20µl of OptMEM medium were added instead of the siRNA mix.
After incubation, cells were lysed with 50µL of supplemented 1X lysis buffer #1 for 30 minutes at RT under gentle shaking, and 16 µL of lysate were transferred into a low volume white microplate before adding 2µL of the HTRF d2 detection reagent and 2µL HTRF Eu-K detection reagent. The HTRF signal was recorded after an ON incubation.
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