2
$\begingroup$

I want to experimentally look at the behavior of antibodies. To do so, I need to be able to bind these antibodies to a substrate.

Eventually, I want to study the specificity of antibodies to know to what substrates they bind. After that, I want to study the biological activity of the antibody over time.

But my question is more basic, namely: to what material do antibodies bind non-specifically? It's not about antigens at this stage, it's just about non-specific binding of antibodies to a carrier material. Also of interest would be howw to get the antibodies of that carrier again.

Improve this question
$\endgroup$
0

2 Answers 2

Reset to default
3
$\begingroup$

Like any other protein, antibodies will aspecifically bind nitrocellulose or PVDF membranes, but any other protein present in your antibody preparation will also do.

Depending on the antibody class, more specific binding can be obtained with protein A or protein G, that recognize the Fc domain. It's usual to have protein A/G immobilized on a stationary phase like agarose.

If you know the antigen used to develop the antibody, you can also use the immunizing peptide crosslinked to CnBr activated sepharose as a probe to bind your antibodies.

Knowing something more about your antibody preparation and what exactly is the 'behavior' your want to study could help obtain better answers.

Improve this answer
$\endgroup$
1
$\begingroup$

96-well plates made from polystyrene are routinely used to immobilize antibodies with excellent results. The immobilized antibodies are then used for binding to antigens e.g. ELISA. The antibody should be free of other proteins for best results as these will compete for binding (many are supplied with carrier proteins such as BSA that will compete).

A good read here:

https://www.corning.com/catalog/cls/documents/application-notes/CLS-DD-AN-454.pdf

Improve this answer
$\endgroup$

You must log in to answer this question.

Not the answer you're looking for? Browse other questions tagged .