You know what assay you need yet it can't be bought off the shelf and you don't have the internal resource to develop it. Why not outsource its development to us? At Imagen Biotech, we generally use the following procedure to develop new assays: (Skip to Flow Diagram)
Literature Search
Before we develop an assay, we check that it is not already described in the literature or commercially available.
Antibody Search
The key to a good assay is a good primary antibody. Unfortunately, commercially available antibodies vary greatly in their quality and
suitability for use in fluorescence microscopy. Therefore, it is necessary to source the required antibody from as many suppliers as possible.
Western Analysis
The antibodies from the various suppliers are then checked using Western Analysis to ensure that they give a clean profile with a band at the right
molecular weight. Any antibodies that give multiple bands or the wrong molecular weight are eliminated at this stage.
Explore Fixation and Permeabilisation Methods
The antibodies are tested in a fluorescence microscopy experiment using several standard permeabilisation conditions to
explore which method gives the best and strongest staining. At this stage the staining profiles of the various antibodies is also checked as they should all be very similar (since they
are binding the same protein). If the staining pattern of a particular antibody does not match the consensus it is eliminated.
Assay Checked for its Functional Discrimination
Since the object of most cell-based assays is to detect changes in the protein of interest, experiments are conducted which
produce a change that the antibody assay should detect. Obviously, these experiments help determine the sensitivity of the assay and the potential signal window the assay possesses.
If Possible, Assay Checked against Null Cell Line
If we have access to a cell line that does not express the protein to which the antibody binds, then the assay is checked
using this cell line to make sure the staining is negative and therefore specific to the protein of interest.
We all understand that assay development is a risky business which is why we will do this type of research at a rate which is comparable to using your own in-house resource (see Pricing.) Furthermore, our staged assay development process gives us the ability to design a research program with lots of No/Go decision points. Should you want to stop the research at any point along the development path then this option is freely available. Likewise, we will keep you always up to date with assay development progress and give you our own estimation of how likely it is that we will develop a functioning assay as we proceed with our development strategy.