I wonder if anyone with experience in the pulse-chase method has compared 35S-Met with Click-iT for labeling of proteins. Which pros and cons regarding to the results did you find and would you recommend Click-iT?
Yes it is very convenient and it works very well. You Can either run a SDS page and UV observation to analyse your proteins or use flow cytometry for a quantitative method. For one gel you must load at least 20 micrograms of proteins which corresponds to 200000 cells. After doing the labeling in the cell culture, i spin the cells and count them so I CAN load equivalent quantities or proteins on a gel. You can stain the gel after UV or blue light observation with compassie to Check the loading. I never used S35 method so I can not compare but click it is probably more expensive. I don.t know neither if it is as sensitive as the radioactive method.I use TAMRA for the gels and an Alexa labeling for the flow cytometry.
Thank you so very much for your response, Marie-line! I have experience from radioactive labeling but not from ClickIt and haven't found anybody who has, so all your comments and tips are very valid to me. :)
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