[3dem] Company Recommendations for Producing Antibodies Raised Against Peptides for IEM

Gina Sosinsky gina at ncmir.ucsd.edu
Fri Jun 27 10:30:24 PDT 2014

Hi Evan,

We used Abgent, a local San Diego company to make polyclonal and monoclonal antibodies (see Cone et al., Frontiers in Pharmacology 2013 for discussions about testing for microscopy). If you don't need to have monoclonal antibodies, then start with polyclonals. They are much cheaper and easier for the company to produce and with monoclonals we had to do a fair amount of interim testing with various clones to determine which was the best. If you have a successful epitope you can always use the same peptide to generate monoclonals at a later stage.  They produced both a monoclonal and a polyclonal for us for the same protein.  Abgent did pretty much everything except for the validation. They had a program that generated a list of unique peptide epitopes ranked according to their predicted antigenicity, made the peptides and then generated the antibodies. There are probably equivalent companies out there, but I liked the fact that they were local and I could easily talk to their chief applications scientist. Other labs here at UCSD have used Bethyl Labs also with success.

We also tried Sigma-Genosys early on, but they were not successful and I wouldn't recommend them.

Is your immuno-EM on purified complexes or in cells or tissue? The former is a much easier because you don't have to worry about antigen accessibility issues like we did in tissue.



Gina Sosinsky, Ph.D.

National Center for Microscopy and Imaging Research
Dept. of Neurosciences
University of California, San Diego
1070 Basic Science Building
9500 Gilman Drive
La Jolla, CA 92093-0608

Office Phone:  858-534-0128
Fax:                   858-534-7497
Lab Phone       858-534-4583
Email :  gsosinsky at ucsd.edu

On Jun 27, 2014, at 8:03 AM, Evan Rossignol <edr5 at bu.edu> wrote:

> Hi all,
> We are starting an ImmunoEM project that requires primary antibodies made against synthetic peptides, a subject I know little about.  We are wondering if anyone had any wisdom or experience with companies to go with or to avoid.
> Optimally they'd produce the peptide and raise the serum and/or monoclonal antibodies.
> Thanks!
> -Evan Rossignol
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> 3dem at ncmir.ucsd.edu
> https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem

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