I would like to check interaction between two proteins. Previously it was reported that 2 proteins I work on interact. I have read a publication where was used BS3 crosslinker. They mapped all Lys NH3 groups and NH3 terminal group in both proteins and compare results when proteins were interacting together.
So I would like to perform exact experiment but of course I have different proteins. I do not have any experience in protein crosslinking. I would be grateful for any help or hints.
Try to get your hands on the book "Bioconjugate Techniques" by Greg T. Hermanson. It is a great reference for crosslinking techniques.
I would first try low concentrations ( > 1mM) of EDC. It is a zero-length crosslinker, meaning it will preferentially crosslink proteins that are close together.
BS3 is a good choice and a popular crosslinker with a medium length spacer arm. It is water soluble so it can crosslink cell surface proteins. A basic protocol can be found here: http://www.proteochem.com/bs3crosslinker100mg-p-39.html
For intracellular protein crosslinking I recommend DSG (http://www.proteochem.com/dsgcrosslinker100mg-p-45.html) or DSS (http://www.proteochem.com/dsscrosslinker100mg-p-74.html). Protocol links are also found in the above linked pages.
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