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8 months ago
jinyu
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10
I find a mAb targeting protein A, which has beed validated to work in wb/IHC/IP or antibody conjugating drugs. I'm wondering if this mAb could be used for blocking the protein A from binding with other proteins? This mAb is the only one antibody which could be bought without having to make it myself, so the answer is really important for me. Looking formard for your responses.
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Thanks for your response. Do you know any other forum where I can post this question?
https://biology.stackexchange.com/
Thanks for helpful response!
Some mAbs are capable of blocking and some aren't. It depends on where they bind. If this Ab is from a vendor, you can check with them to see if it blocks.
Really helpful response! Yes, the Ab do comes from a cendor, but they haven't validate it's blocking effect. Is there any difference between blocking mAb and other mAbs. In my opinion, when mAb binds to the protein A, the other Abs are hard to bind again. So every mAb is able to function in blocking , where the only difference is their strength of the blocking effect.
Protein A binds the Fc portion of antibodies, whereas it sounds like you have a mAb whose variable region binds to protein A. I've typically used protein A for antibody purification, and I'm not sure what your application is. That said, I wouldn't assume that your mAb is blocking. In order for it to block, it would have to bind the antibody-binding site of protein A in a way that prevents any further binding and it would have to do this in a saturating way (i.e. you would need to use enough antibody to bind all of the protein A molecules so that there wouldn't be any free protein A in solution). In theory, your mAb could bind a different epitope from the protein A antibody-binding site, which would mean that it isn't blocking. Maybe you could devise an experiment to test its ability to block (e.g. use ELISA to see if your mAb can prevent protein A from binding some other antibody from a different species).
By the way, if you're still looking for another forum, you could try ResearchGate:
ResearchGate
Thanks for your suggestion again! Wether just taking a flow cytometry staining or ELISA as you suggest is enough to verify the blocking effect when conducting scientific research. Is there any other experiments I have to do?
A well-designed ELISA with the proper controls would be pretty convincing, but it would be nice to confirm blocking with a second technique. If you have a supervisor or PI, you could check with him or her.