[Xenopus] Antibody update

[Dominique Alfandari]

Dear Colleagues, 
we have now been working to make monoclonal antibody to Xenopus proteins for one year. I wanted to update you on what we have and what we are doing now.
We have screened by immunoprecipitation more than 400 clones from mice immunized with cytoplasmic and membrane proteins isolated from tailbud stage Xenopus embryos. We have identified more than 50 antibodies that efficiently IP proteins of different sizes. We are grouping them in Size groups and will start the final characterization by mass spec in the coming month. We have about 1000 more to screen.

As for targets:
We have mAb to Ror2, Par3, Rax1, Lbh, PCNS
We have splenocytes for Xbra, Smad7, Sox3, Ror2, Rax
We have mice immunized with Ngn2, Slug, Sox8, Twist, Prdm12, Sox9, FoxD3, NeuroD.

The antibody all work by IP and IF on transfected cells, most work by IP on embryo extracts. Some are being characterized in labs that originally requested them. As soon as we are satisfied with the characterization we will make them available and write a descriptive sheet for all of them. If you want some sup to test yourself, contact me with you FEDEX number.

We just invested in an electrofusion system which appears to be magnitudes more efficient than our traditional PEG in our first experiment. From Frozen splenocites, we obtain 7 fold greater number of hybridoma than with fresh spleen with PEG. The proportion of hybridoma producing IgG to the targets is also much greater.

Dom
Dominique Alfandari
PhD. Professor 
alfandar at vasci.umass.edu <mailto:alfandar at vasci.umass.edu>
661 north pleasant street,
ISB rm 427B, Amherst, MA, 01003



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