Dear colleagues,
I wanted to measure antibodies concentration by indirect ELISA . I have coating my microplates by control antigen and modified antigen . I have the absorbances but I have a problem with calibration curve ( it is incorrect) . I can't also do qualitative dosage because the absorbance of 2nd standard is incorrect. I have only the problem with standards. Is there any method or formula to compare Absorbance of control antigen and modified antigen without calibration curve ? to only say if the immunoreactivity is increasing or decreasing.
Thank you
Dear Hamida,
if one of your standard is wrong, just move out its absorbance from the curve.
About your question, it is possible to know if your immunoreactivity was affected in your modified antigen. Just take the mean absorbance value (considering dilution factors of course) of your unmodified antigen as 100% and compare with the mean absorbance value of your modified antigen.
Dear Ana Maria,
thank you so much for your answer . Unfortunately ,all my standards are wrong , probably its denatured. Please do you have reference how use only absorbance to measure immunoreactivity ? ABsorbance of antigen control 100% and absorbance of modified antigen ?.
thank you so much
Dear Hamid
As I see there is quite differences between the antigen and the modified antigen in the standard curve calibration ...It may due to the size of modification to the antigen..Also you have to know wether antigen have been denatured or no, also
Dear Mr Ziad ,
standards curve was made with control antigen only . to see if modification of antigen increase or decrease immunoreactivity. The problem was in my standards ( antibodies ) . I want to exploit my results because they are good.
thank you
Dear Hamida,
A file is attached where immunoreactivity was made with percentage.
Good luck!
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