Im working on a protein. We have our cell line and we suspect that the protein is present there. However, we don't have any commercial antibody for this protein so we cannot do any western blot to check. We can detect the protein at the mRNA level and is relatively abundant.
So my question is, how do I know for sure that my protein is there? Like how do I know if the mRNA is actually translated into actual protein? Is there any technique other than western blot to confirm this?
Thank you very much
What is your pro and its function?
On way is you check the function.
Other way you can pure it with gel or hplc and so compare with a standard or do sequence.
This was my idea
The function of the protein is unclear. It's difficult to observe the phenotype though, it's not always black and white. Thats why I need a more robust method to know if the protein is really there.
Good question.
Thus, we must have to develope the new quantitative proteomics PDMD (Protein-Direct-Microsequencing-Deciphering) method (please see files; HepG2 fucoidan and JMBT alopecia).
By the way, HPLC-MS, ELISA, PCR and Western blotting are not quantitative methods at all.
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