How to test heat inactivated Normal Human Serum (View forum version)



CMIRC

Posted 24 April 2013 - 07:36 AM

Dear All

I need some help. I am working with complement protein iC3b and need to heat inactivate Normal Human Serum (NHS) to inactivate iC3b as a control. However, it is not working. At the moment I heat NHS at 56 degrees for 60 min and then freeze it for storage in -80 freezer. Whenever I use it for complement assays and label cells with the complement antibody I get binding. This therefore means that the NHS has not been inactivated totally. I have tried the conventional method of heating at 56 degrees for 30 min but even that did not work. I would therefore like to find out a quick way to test if the NHS has been heat inactivated.

I am open to suggestions.

Curtis

Posted 25 April 2013 - 11:03 PM

a friend of mine had the same problem last time with heat inactivation. above 58C he got solid phase. then I think he increased temp from 56 to 57 in hot block (not water bath) and then tapped every 5-10 min to make sure the serum mixed.