In article <[email protected]>, [email protected] says... > > Hello, > > We are a group of Berkeley MS students in the Chemical Engineering > department. We are currently engaged in a research project aimed at > evaluating current trends and drawbacks of the extant technologies used in > the field of proteomics today. In particular, we are interested in Western > Blotting and ELISA assays as well as the potential improvements/replacement > assays that are currently being developed today. We were wondering if it > would be possible for us to set up an interview with you sometime next week > to get your opinions and expertise in the field. In addition, if you do > not personally have any experience in the field, could you refer us to > anyone contacts you may have either in academia or industry who has > relevant experience? Any help you could provide would be much appreciated. > > Thank You, > Brian Kim and Rishabh Jain
A very simple improvement for ELISA are dot-blots. The sample is sucked through a PVDF-membrane (eg, in a 96- well filtration manifold), proteins bind quantitatively to the membrane. This is then developed with peroxidase- or phosphatase-coupled antibodies, detection is by chemiluminescence counting. This results in a very sensitive assay with a linear range of 5-6 orders of magnitude (ELISA about 1 order). The main advantage over ELISA is that binding to plates in ELISA is very inefficient, only a few % of the sample molecules actually bind to th plastic and can be detected. PVDF-membranes bind > 95% of proteins present in the sample, thus you get an increase in sensitivity by almost 2 orders of magnitude. The limitation of dot-blots is that it cannot be used to detect small molecules (haptens). -- Car: Erratically moving obstacle on the road _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
