[Xenopus] Antibody grant

Dominique Alfandari alfandar at vasci.umass.edu
Fri Apr 23 08:46:27 EDT 2021

Dear Colleagues,
My R24 to make monoclonals for Xenopus protein is running out this summer. During these 4 years, we have produced, subcloned and partially characterized over 120 hybridoma lines. I am thinking about resubmitting a new application that would be more straight forward. I plan to only do target proteins instead of crude embryo extracts given the relative success of both approaches. My plan would be to continue down the list of request, using fusion proteins for cytoplasmic and nuclear proteins and mRNA vaccine for transmembrane and secreted proteins. I will be testing this last approach over the next few month to see if it is viable for our system. Another goal is to produce mAb to the usual abundant contaminant (yolk for example), so that we may produce a cocktail that can be used to preclear IP/co-IP prior to mass spec and increase the number of relevant proteins detected. DSHB could produce this cocktail and distribute it as a single sup possibly.

This email has 3 goals: 1 see if there is interest/support from the community (If yes please send me emails so that I can document this in my resubmission. 2 See if any lab would like to partner as a secondary testing site. So far Carole Labonne' lab has done a fantastic job, but ideally I would need more people willing to do this. At least the people that have requested the antibody have to be ready to test them in their system. 3 To see if it might be of interest to include Axolotl and other model organism in this production.

IF you have not followed our progress (Sorry I have not been too good at this), we have produced some great antibodies to transcription factors (Xbra, VegT, Sox8, Sox9, Sox3..) as well as other cytoplasmic and secreted proteins including Wtip, Rhor2, Par3. Recently one of our by product an Anti Flag was advertised by the DSHB. We have also made a great antibody to Zs-Green that works in IF and FACS. We have much more antibody that have been characterized on the target but have not been validated in embryos (Ngn2, Ascl1, Ets1, Zic1).

Please let me know what you think.

Dominique Alfandari PhD
Preferred Pronouns: He/Him/His
Professor, UMass Amherst
ISB 427B, 661 North Pleasant street
Amherst, MA 01003.
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