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I have a question regarding one of the step in the protocol. Step (h) in the sample preparation says the “if cGMP concentrations in your samples are very low, the acetylation reagents can be dried after step h, without dilution step i to minimize the volume. Then reconstitute in a 50-100ul volume of Assay Buffer”
We are experiencing difficulties detecting cGMP levels in our sample of THP1 cells. Can I ask how to “dry” the acetylation reagents in step h?
Asked on Nov 08 2013
After adding the acetylating reagent mix, incubate at RT for 10 mins, then put the tubes in a laminar airflow hood or simply air-dry. 5ul will dry-up fairly quickly. For low amounts of cGMP, add 50-100ul of the assay buffer to dried tubes and then proceed with quantification.
Answered on Nov 08 2013