Hey!
I am developing an Elisa against anti-GRP78-antibodies for measuring titers in serum samples. I want to coat the plate with GRP78 in a carbonate buffer and detect the auto antibodies with anti-human-igg/igm/iga antibodies. As a standard row, i want to apply different amounts of murine anti-grp78-antibodies on the plate after coating and detect them with anti-mouse-igg. Does this standard row make sense? I am unsure, because i use different detection antibodies with probably different properties. I hope that you understand my question.
Best regards, Aaron
It would be better to use the same antigen and antibody for the standard as for the samples, i.e. human.
I think you will have a problem with fitting curve and quantification of results after.
Adam B Shapiro Yes i know, but i dont have purified human anti grp78. I have a serum from a patient who is known as anti grp78 positive, but there is so quantification.
I wouldn't recommend using a different species because like you said, you would have to use different antibodies that would bind at a different specificity. I would try to find a recombinant protien of the same species in a known concentration as your standard. Keep in mind that most recombinants are only a certian length of the sequence so you have to make sure your antibody binds to that part of the sequence.
- Badges
- Science method
- Similar topics
- Medicine
- Immunology
- Immunology Techniques
- Immunoassay
- ELISA