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Serum Batch Test....


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#1 molecular_medicine

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Posted 20 April 2009 - 10:33 AM

Hi all,

I am in desperate need to do a batch test for the serum (FBS) we have. We usually heat inactivate the serum and then use it for our cell culture. I please need your help with the following:

1) why do we do heat inactivation?

2) if I need to do a serum batch test for a particular protein, is it ok I either do an ELISA or western blotting??

Also, I am not able to find an antibody against bovine for the protein that I am interested in. Not able to find an ELISA kit as well. Although, antibody for rat, mouse and human are available in plenty (both antibodies and elisa kits)..

Please suggest what I need to do. Thanks.

#2 bob1

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Posted 20 April 2009 - 04:46 PM

Heat inactivation stops the complement cascade lysing the cells.

I would probably do ELISA for a particular protein as western isn't sensitive or reproducible enough.

#3 molecular_medicine

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Posted 21 April 2009 - 01:43 AM

Heat inactivation stops the complement cascade lysing the cells.

I would probably do ELISA for a particular protein as western isn't sensitive or reproducible enough.



Thanks a lot for the suggestion... will update on the progress..... :)

#4 sgt4boston

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Posted 22 April 2009 - 08:36 AM

If you do the elisa you should run in parallel with the dose response curve a curve of the purified protein (bovine?) that you are quantifying. The binding may be different between species. If the curves (in the same matrix) are identical the results can be directly interpreted. If not you will have to read the results from the purified protein curve you run.




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