Which reagents do you currently support for making affinity capture screening matrices?

Our currently supported reagent list can be found here.

What is the importance of the 'Single Reagent Flag,' 'Elution Method,' and 'Elution Reagent' fields?

Our system is designed to cross-reference the in vitro solutions used in affinity capture with the analytical results obtained in the form of SDS-PAGE banding profiles and e.g. mass spectrometry. The nature of the solutions used, and the number of different solutions used, will affect the result. Our aim is to help users of our system find the best conditions of affinity capture for their application – and we aim to understand the solution parameters affecting the behavior of protein complexes in vitro.

In many cases excellent results can be obtained using a single step affinity capture with a single extraction and washing solution. Please refer to Hakhverdyan et al. 2015. However, if your results were obtained using multiple solutions, you can still benefit from the tools available at copurification.org. In this case, please indicate 'no' in the 'Single Reagent Flag' field of the Sample Descriptions File and add details in the Notes section. We also suggest you upload an MIAPE compliant document of the experimental procedure.

Likewise, results obtained by SDS-PAGE and/or MS may be affected or biased by the method of elution from the affinity medium. Some elution solutions are not powerful enough to uniformly release the affinity immobilized protein complexes. We recommend a solution containing 2% w/v SDS as the solution of choice for reproducible and uniform elution. Please indicate if denaturing or native elution was used in the 'Elution Method' field of the Sample Descriptions File, and further indicate the specific elution reagent and solution in the 'Elution Reagent' field. All the above are further described in the HOWTO.