I am analyzing the levels of secreted HBsAg in the hepg2.215 culture supernatant by commercial enzyme immunoassays (Monolisa™ HBs Ag ULTRA). My questions are:
1) Can i perform the test in a 96 well plate?
2) What is the incubation period required before taking the supernatant?
3) What is the minimum number of cells that can be used (seeded) in the well plate?
We have been using the PLC/PRF-5 for doing the HBsAg immunoassays. We have observed a peak in HbsAg levels in the supernatant on day 4/5. This is ideally when we assay the cell free medium.
Just one question: Does any one have the HepG2.2.15 that can be shared with my lab?
Regards.
Hi Arpit Shah ·As you know, the above protocol is measure HBsAg in serum, not in cell culture, last week i carry the test as follow: cells were seeded in 96 well plate. on day three i took the supernatant and do the test steps according to the protocol, but no result (except the positive control).> why??
Dear Ahmed,
You'll need to use HbsAg ELISA kits that are very sensitive, and are capable of measuring the antigen from cell free supernatant. This works in our labs.
Regards.
Hi All, I am using Biorad ELISA kits (Monolisa™ HBs Ag ULTRA) to measure antigen in serum free medium, but no result ( 96, 48 and 12 well plate,). cell free supernatant from T 75 culture flask we detect antigens using the same kits.
Best Regards. Should i shift to more sensitive kits from other company ?
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