Hello,
I bought this antibody to make electrochemical immunosensors, I bought the ab93903 antigen which is recommended. I prepared the antibody to the desired concentration in 10mM PBS pH 7.4 and tested various antibody concentrations. I also dissolved the antigen stock (ab93903) in 0.1% acetic acid, which is also recommended, and then prepared the standards in 10mM PBS pH 7.4. But the sensors are not responding to the standards, am I doing something wrong? Do you have any recommendations?
Thanks.
Thanks Wolfgang Schechinger for your answer.
I funcionalize the SPE with COOH or NH2 and then I active the COOH groups in SPE or in antibodies (in case of SPE funcionalized with NH2) with EDC/NHS to attach the antibodies.
When you add EDC to the antibodies, you're very likely crosslinking the antibodies to themselves, as they have plenty of accessible amino groups (Lys, N-termini), so that won't work
Activating the sensor surface this way should work.
Could you give some details on the coupling protocol, maybe?
Is there any way to check if the antibody has bound? E.g. a changing resonance frequency, impedance/condictivity? What is the detection principle?
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