<html><head><meta http-equiv="Content-Type" content="text/html; charset=us-ascii"></head><body style="word-wrap: break-word; -webkit-nbsp-mode: space; line-break: after-white-space;" class="">Dear Colleagues, <div class="">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.</div><div class="">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.</div><div class=""><br class=""></div><div class="">As for targets:</div><div class="">We have mAb to Ror2, Par3, Rax1, Lbh, PCNS</div><div class="">We have splenocytes for Xbra, Smad7, Sox3, Ror2, Rax</div><div class="">We have mice immunized with Ngn2, Slug, Sox8, Twist, Prdm12, Sox9, FoxD3, NeuroD.</div><div class=""><br class=""></div><div class="">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.</div><div class=""><br class=""></div><div class="">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.</div><div class=""><br class=""></div><div class="">Dom<br class=""><div class="">
<div style="color: rgb(0, 0, 0); letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space;" class=""><div style="color: rgb(0, 0, 0); letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px; word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space;" class=""><div style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space;" class=""><div style="color: rgb(0, 0, 0); font-family: Times; font-size: 14px; font-style: normal; font-variant-caps: normal; font-weight: normal; letter-spacing: normal; text-align: start; text-indent: 0px; text-transform: none; white-space: normal; word-spacing: 0px; -webkit-text-stroke-width: 0px;"><div><b class=""><i class="">Dominique Alfandari</i></b></div><div><b class=""><i class="">PhD. Professor </i></b></div><div><b class=""><i class=""><a href="mailto:alfandar@vasci.umass.edu" class="">alfandar@vasci.umass.edu</a></i></b></div><div><div class="">661 north pleasant street,</div><div class="">ISB rm 427B, Amherst, MA, 01003</div><div class=""><br class=""></div></div></div></div></div></div><br class="Apple-interchange-newline">
</div>
<br class=""></div></body></html>