UCSF was chosen as a beta test site for the Cell Biosciences Firefly. The Firefly is an automated western analysis robot. Cell lysate or protein samples are first separated according to their pI using capillary electrophoresis. Subsequently, the protein of interest is quantified using antibody-mediated detection. Separation of the proteins by pI allows analysis of the protein of interest and all of its charged post-translational modifications using only one antibody.
This instrument is particularly useful for analyzing proteins thought to have charged modifications and/or where the sample quantity is limited. In addition, because the Firefly performs automated analysis of a 96-well plate in 8 hours, this instrument is also useful for medium-throughput applications.
You must be <trained> by a member of the Core Facility before you schedule or begin using the Firefly.
When you contact the facility, please have the answers to these questions available:
- Protein: What protein(s) are you interested in studying? Are they tagged? Are they membrane, cytosolic, or nuclear (or other)? Do you know the pI (calculated or measured)? Are there post-translational modifications you are interested in? Have you studied this with IEF before, either in 1D or 2D gels?
- Lysis buffer: Can you try preparing the sample using the lysis buffer shown below?
- Antibodies: What antibodies do you typically use on a western? What secondary is required? What dilutions do you use? Do you have other antibodies to this target available? We do not always find that the "best" antibody in a western is the best in our Firefly system, so it can be helpful to try several in the first runs.
We do provide all disposables for general use.
Location - GH-S252
- Charge
- UC - no charge
- Non-UC Academic - no charge
- Non-academic - not available
