Very high background in ELISA
Posted 26 September 2010 - 09:42 AM
Posted 27 September 2010 - 02:56 AM
If the patient's samples are reacting you can pre-incubate them with the blocking solution before testing. Can you dilute your sample to any degree..say 1:10?
Do you have surfactant in the wash solution? You can also allow the wash to sit in the wells for 1 minute before decanting/blotting.
Lastly, and I don't think it applies at this time are 'heterophile abs' in sample.
Posted 27 September 2010 - 06:23 AM
Edited by shilpeejnc, 27 September 2010 - 06:28 AM.
Posted 27 September 2010 - 11:24 AM
2. As you are diluting the patient's samples...1:100 which is good and using some fcs. Incubate the sample and diluent one hour is fine. The purpose being to adsorb out any interference. You could also use the same animal type as the secondary ab( conjugate) which could be goat or rabbit etc or mix all three.
3. I don't understand your question for incubation of secondary ab. this could be 15 min to one hour.
4. Can you describe your coating/blocking steps?
Posted 28 September 2010 - 09:43 PM
Posted 29 September 2010 - 02:37 AM
Posted 29 September 2010 - 06:57 PM
for negative I take HIV negative sera. for positive I dont take any thing sepsrately. But why this question?
Do you have any positive 'control' sera to test. Known positives; or any commercial control material. Your test is to screen samples but what is serving as your positive and negatives?