Neutralizing 5% TCA - Neutralizing TCA for use in cAMP Enzymeimmunoassay (May/11/2011 )
Hello there,
When I google search "neutralizing TCA" all I get is stuff about baking soda for cosmetic chemical peels or info about tattoo removal.
I'm using neuronal cultures. I need to measure their cAMP production after they were exposed to various treatments. We're going to assay both the NB they were in with treatment, and the 5% TCA they are now in. The cAMP EIA kit from GE Healthcare comes with instructions to neutralize TCA with diethyl ether. My lab manager would prefer a less volatile approach if possible. I thought about 1 M Tris buffer but I wonder if that may interfere with the immunoprecipitation process. Any other ideas?
Thanks.
-nueropharm-
nueropharm on Wed May 11 19:12:52 2011 said:
Hello there,
When I google search "neutralizing TCA" all I get is stuff about baking soda for cosmetic chemical peels or info about tattoo removal.
I'm using neuronal cultures. I need to measure their cAMP production after they were exposed to various treatments. We're going to assay both the NB they were in with treatment, and the 5% TCA they are now in. The cAMP EIA kit from GE Healthcare comes with instructions to neutralize TCA with diethyl ether. My lab manager would prefer a less volatile approach if possible. I thought about 1 M Tris buffer but I wonder if that may interfere with the immunoprecipitation process. Any other ideas?
Thanks.
Hola,Tris is a biological buffer that is in the storage medium of the majority of pure proteins for activity assay, in order to add less volume possible I woul prepare a Tris base solution wich has a pH of 12 or 13. Other possibility is any alcali as NaOH. . according to the sample volume that you have if itīs very little I would assay the neutralization in big volume in a pHmeter and after reduce the neutarlization agent proportionaly to the relationship betwen both volumes: pHmetr/sample. Buena suerte
-protolder-