Dear friends, I would like to carry out a screening of drugs that induce the expression of glysosylated proteins. I would treat fibroblasts, and reveal the specific proteins with primary antibodies and secondary flueresceinated antibodies, or primary fluoresceinated antibodies. can you suggest protocols? Can you suggest papers that Icould read? thank you very much ANNA
Hello and welcome!
You might begin reading review articles on high content screening to get an overview.
When you’re ready to dig into details, check out the amazing NIH Assay Guidance Manual: https://www.ncbi.nlm.nih.gov/books/NBK53196/toc/?report=reader#!po=3.57143
can you suggest some papers? our target is measuring the total amount of
fluorescence of a target protein in cells treated and not treated with a
You might begin by taking a look at @Christian_Tischer posts in this related thread about measuring total amount of a protein’s fluorescence: When is it recommended to normalize protein fluorescence to nuclear fluorescence?
A lot of papers take the approach of measuring mean or integrated intensity of fluorescence signals but most will not describe the theory behind it as well as Christian does.