We are putting ELISA for tissue samples. the protein amount which we coat at ELISA plate was 100 ng which is total extracted protein. So we take primary antibody and secondary and interpret results. we are using 12 different antibodies for one sample for one protein 12 reaction we put. But not having any standard antigen. So can we determine the amount of that particular 12 proteins from one protein samples without standard antigen?
No - you cannot estimate the amount of target protein without using standard.
What you can do is to estimate possible difference in the protein amount between two different samples (e.g. before and after treatment) per equal concentration of total protein .
Depending on how many samples you have (and therefore how many plates you use) you cannot even properly estimate the differences in one protein if you have samples on more than one plate. Which proteins are you testing for? Is there any way for you to purify or buy them for use in standards? ELISAs without standards are only useful to show that your antibody binds to something, but you cannot even show that actually is specific for your protein.
Without a standard you can not quantify your target proteins. You have to use dilution series ( e.g. 0, 20, 40, 60, 80, 100 ng) of well quantified standards and compare the readings with the respective target proteins.
As you did not determine, nevertheless the concentration of each protein will be approximate. We determine protein concentration, using a protein electrophoresis on one standard.
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