I'm using 19C7 2ug/mL as capture, and M18 1 ug/mL as detection for Troponin I in ELISA, with SA-HRP.
somedays, I get reliable results, and many others, it's all over the place :(( when contamination happens, I get 1.2 reading for all the wells, including the blank.
Then I have to prepare new reagents every few days, less than a week.
It's very frustrating, how can I identify the contaminating agent?
It could be due to insufficient washing. You could check if the ports on the washer are blocked.
take care of the following-
check pH of PBS and make sure it is not contaminated. reagents except PBS should be freshly prepared- not an issue. You should increase the number of washings and take care that the ELISA plates are properly tapped. give 4 washings after blocking step and 6-7 washings thereafter for each step.
Before test all solutions for ELISA should be fresh and spinned (10 min at 10 000 rpm). Usually we use 5 washings after each step. Washing solution contains 0,05% of Tween 20; wells should be filled completely.
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA