I am using 5% HSA (human serum albumine that we make) to make a standard curve for ELISA. I spike it with either SPA or AFP or any other antigen then use the prizm software to find the concentration and to make a standard curve. We usually compare the new standard with the old lot and are trying to make the BioRad within the range, the range that has been puplished by other companies. Is there any way to know my standard concentration other than comparing it with the previous lot or with a standard curve from another company? Also, if my BioRad out of the range (higher or lower) does switching to another range from another company make a difference?
Can you determine the concentration of your protein before you spike it into your standard HSA mix (if it is pure then you can use a simple bradford/BSA etc) and then add enough to get the concentration you want?
Is 5% HSA solvent for dilution of quantified antigen (AFP etc)? Did I right undestand You?
I think You should use sertified antigen standard or measure its concentration yourself by suitable method (Lowry, Bradford, HPLC etc) BUT NOT ELISA.
What is BioRad ? Is it some reader for ELISA from BioRad?
If your sample is out of range, You should find suitable dilution.
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