I wanna analysis cytokines level in different stages of CIN and ca cervix both in HPV DNA positive and negative cases. We collected 1000microliter cervical swab by cyto brush for detection of HPV DNA by hybrid capture. We processed this sample by adding 500microL denaturating agent. We have some leftover samples after hybrid capture..
Is it possible to use that remaining sample to analyze cytokines level or I have to recollect cervical swab? If I have to then how to collect, process and preserve the sample for both cytokine detection by flow cytometry and hpv dna and mRNA by PCR..
Hi Paroma, I certainly think its possible. You would certainly need to employ protease inhibitors up front (complete from SIAL is a good one) and assess the tissue protein level using say Pierce BCA proteein assay kit. Elisa kits for cytokine could then be employed but you would probably need the highly sensitive variety due to the small amounts of tissue. Bead array analysis on FCM might work for you in place of ELISA. The mRNA side I can't comment on.
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