I would like to measure the function of CFH as a decay accelerator, but all the protocols I am reading are using a machine to do so. Is there a way to do it by hand? Possibly just mixing together all the necessary components and running an SDS-PAGE gel for silverstain/coomassie or possibly a western to look for decay?
Anything will help- thanks!
You can do it manually using C5-depleted serum and guinea pig serum. Detailed description of the procedure you may find in our paper: http://www.ncbi.nlm.nih.gov/pubmed/23071769
The advantage of this method over the others is that you assess decay accelerating activity in the physiological milieu of whole serum. Most of the methods use purified components and so one can miss some relevant interactions between serum components or augment something, which does not play a role in physiological conditions.
- Badges
- Science method