I need to know if there's a way I can specifically quantify (visually) his-tag fusion protein. I have checked for commercial his-tag stain reagents, but I'm looking for state-of-art method(s). Please send your suggestions.
Hi Saaraj. I'm not quite sure what do you mean by "quantify (visually)". Your question is not clear. In which platform you would like to view. Is your protein is the extracted protein, OR you would like to visualize your protein in cell culture based system?
If it is the extracted protein, I guess you should run the protein on SDS-PAGE and perform Western Blot. Then, you can stain the membrane using the commercially available Ab. There are software the one that I know is Quantity One (Biorad) may allow you to do the analysis to quantify your protein band by scanning the blot and do the analysis.
Thanks for your suggestion. Protein will be in cell culture based system when I want to quantify it.
If you want to quantify in cell culture system, provided that you have the antibody which tagged with fluorescence, then, you can do the cell staining. But you must also stain the nucleus to allow you to count the total number of cells. (His-Tag labelled cells/ Total no of cells).Usually, for this case, we use florescence microscope. It is quite tedious, but sometimes, it is better that way if the expression of you protein of interest is not high enough, which may cause difficulty to observe by Western Blot.
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