tag:blogger.com,1999:blog-3693238014525650573.post1612329094211799589..comments2024-03-28T08:46:55.598-07:00Comments on News in Proteomics Research: Why do your alkylation and reduction separately? Do them together! Unknownnoreply@blogger.comBlogger6125tag:blogger.com,1999:blog-3693238014525650573.post-66230222265348327342020-09-18T05:33:42.903-07:002020-09-18T05:33:42.903-07:00This comment has been removed by a blog administrator.MKniermanhttps://www.blogger.com/profile/06471635276260004397noreply@blogger.comtag:blogger.com,1999:blog-3693238014525650573.post-36745224682736517062018-03-07T08:27:56.554-08:002018-03-07T08:27:56.554-08:00I'm curious with that as well Rob. After comin...I'm curious with that as well Rob. After coming across this several weeks ago, my first two attempts utilizing the CAA/TCEP combo resulted in only 75% labeling efficiency. As Chris mentioned, further testing is always needed to see what can be achieved in your hands.Anonymoushttps://www.blogger.com/profile/05876605388465473455noreply@blogger.comtag:blogger.com,1999:blog-3693238014525650573.post-15261283625017474962018-03-01T06:16:57.306-08:002018-03-01T06:16:57.306-08:00I'll be impressed if this is now the only meth...I'll be impressed if this is now the only method the Mann group and their many academic descendants use... is it?Rob Beynonhttps://www.blogger.com/profile/16898940172516762128noreply@blogger.comtag:blogger.com,1999:blog-3693238014525650573.post-47099619705630465062018-03-01T01:55:27.786-08:002018-03-01T01:55:27.786-08:00Extensive and quite recent exploration of IAM vers...Extensive and quite recent exploration of IAM versus CA here : https://pubs.acs.org/doi/10.1021/acs.jproteome.7b00022anohttps://www.blogger.com/profile/00219579136338577888noreply@blogger.comtag:blogger.com,1999:blog-3693238014525650573.post-24421006793596175182018-02-28T13:41:38.810-08:002018-02-28T13:41:38.810-08:00I've been using iodoacetamide/TCEP at the same...I've been using iodoacetamide/TCEP at the same time for several years. I'll have to revisit the idea of using chloroacetamide.Anonymoushttps://www.blogger.com/profile/13866994858092347096noreply@blogger.comtag:blogger.com,1999:blog-3693238014525650573.post-55019335756848232242018-02-28T07:05:21.241-08:002018-02-28T07:05:21.241-08:00Interesting post, but I would suggest critically r...Interesting post, but I would suggest critically reading the paper you link above in MCP regarding off-site modifications with iodine containing reagents. There is a lot of interesting information contained within it regarding other reduction/alkylation combos, and you may not conclude that TCEP/CAA is the best choice after reading it. There are also other things to consider, such as the composition of your lysis solution, which if you are using deoxycholate, TCEP will likely not be the best choice for you (TCEP will precipitate deoxycholate). In addition, one aspect that is very clear in the above paper is that the actual method that you use for your processing can make a much larger difference than the reagents used. The observations of that work were strongly apparent in the in-gel data, but far less so in the in-solution (FASP-treated) samples (for reasons discussed in that work). This is also just the results of one lab processing in their hands...I wonder what the result would be looking for these unexpected modifications in a much larger data set (perhaps in the MSFragger paper, or the wide search window Gygi lab paper)?<br /><br />I'm not trying to advocate for the use of IAA (or any other reagents), but rather to suggest testing and tailoring your workflow to the results you obtain in your own hands. <br /><br />Cheers,<br />ChrisAnonymoushttps://www.blogger.com/profile/00615323186043032713noreply@blogger.com