Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Protocols and Methods Forum
Register Search Today's Posts Mark Forums Read

Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.

embryo powder

embryo powder - Protocols and Methods Forum

embryo powder - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.

LinkBack Thread Tools Display Modes
Old 03-07-2008, 08:19 PM
Ed Siefker
Posts: n/a
Default embryo powder

We're going to be making some embryo powder for antibody preabsorption
for in situ hybridization. Pretty much as per this protocol:
[Only registered users see links. ]

We're wondering whether it's ok to pulverize the embryos in liquid nitrogen
before homogenization in PBS. I'm not sure our homogenizer would be able
to handle whole embryos. That's not going to affect the antigenicity of the
embryo powder, is it?

Reply With Quote
Old 03-08-2008, 06:20 PM
Posts: n/a
Default embryo powder

In article <[Only registered users see links. ].net >, Ed Siefker <[Only registered users see links. ]> wrote:

That would be totally fine.

I have one problem with the protocol though:
Some/many proteins from aceton powder redissolve when
you hydrate it back. So they will end up in your antibody solution
and the antibody against them won't be preabsorbed at all.

Although this will provide some competition with the
antigens in situ, there still will be, inevitably, some

If I were doing it, I would make a who protein extract from
the embryo and coupled the total soluble protein fraction
to a CNBr-activated agarose. Then preabsorb with aceton powder
thoroughly *washed* in buffer (this removes Ab against
insoluble antigens), after which incubating the Ab with the
immobilized total protein will remove Ab recognizing soluble
antigens. More work but better results.

Reply With Quote

embryo , powder

Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
baking powder and bones Keith Michaels Botany Forum 7 11-03-2010 06:05 AM
embryo decrease James Zebrafish Forum 0 04-15-2008 04:02 PM
[Zbrafish] Mass Embryo Rearing christian lawrence Zebrafish Forum 0 05-03-2007 05:38 PM
[Zbrafish] embryo contamination christian lawrence Zebrafish Forum 0 04-05-2007 12:15 PM
Noncharged Cellulose Powder cryobeer Protocols and Methods Forum 0 05-10-2005 02:20 PM

All times are GMT. The time now is 02:13 AM.

Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2015, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.11984 seconds with 16 queries