Comparison of Two Methods for Screening TCR Vβ Biases of T1DM Patients: Silver Staining for Polyacrylamide and Ethidium Bromide Staining for Agarose Gel Electrophoresis
Objective: To explore a fast and sensitive method for screening T cell receptor (TCR) biases through comparison of silver staining for polyacrylamide gel electrophoresis and ethidium bromide staining for agarose gel electrophoresis. Methods: The lymphocytes were isolated from peripheral blood which collected from ten patients with T1DM; the total RNA was extracted followed by transformed to cDNA, and then electrophoresis was carried out on polyacrylamide and agarose gel simultaneously, finally stained with silver and ethidium bromide dying respectively. Results: The pictures of TCR Vβ biases detected with silver staining for polyacrylamide gel electrophoresis were very similar to that with ethidium brominde staining for agarose gel electrophoresis, Vβ7, Vβ11 and Vβ17 were the mainly biased genes with higher frequencies, 10 and Vβ13.2 were the common genes which did not biased in all the ten patients. In some Vβ genes, the number of the bands detected with the former technique was more than that with the latter methods. So did the clarity of some bands. Conclusions: Silver staining for polyacrylamide gel electrophoresis is a fast, sensitive and safe method for screening the biases of TCR Vβ genes.