Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Animal and Molecular Model Systems > Arabidopsis and Plant Biology
Register Search Today's Posts Mark Forums Read

Arabidopsis and Plant Biology Discuss Arabidopsis and Plant Biology Research.

Protoplast transformation and HA antibody.

Protoplast transformation and HA antibody. - Arabidopsis and Plant Biology

Protoplast transformation and HA antibody. - Discuss Arabidopsis and Plant Biology Research.

LinkBack Thread Tools Display Modes
Old 01-31-2006, 06:58 PM
Hicham Zegzouti
Posts: n/a
Default Protoplast transformation and HA antibody.

Dear Arabidopsis community,

I am trying to express an HA-tagged protein (50KD) in protoplast
using PEG transformation. When I run a western after 18-24 hours
incubation, I don't see any differentially present band corresponding
to 50 KD present in my transformed protoplast lane. All the bands
that were present on the gel are detected non specifically by the HA
antibody I am using (Covance). When I use GFP as a control for
transformation, the GFP antibody recognizes the GFP band showing that
the transformation protocol I am using works at least for a 30KD
protein. I am thinking that the HA antibody (Covance) I am using may
not be good.

I would be very greatful if you could tell me about any experience
you have with HA tagged proteins and what antibody you have used
successfully for westerns and immunoprecipitations from plant, plant
cells or protoplasts.

Thank you from your help

Hicham Zegzouti, PhD

************************************************** ************************
Department of Molecular, Cell and Developmental Biology
Christensen lab
621 Charles E. Young Drive, South
P.O. Box 951606
Los Angeles, CA 90095-1606
Phone/Fax: 1 310 794 7096
email: [Only registered users see links. ]
************************************************** ************************

Reply With Quote

antibody , protoplast , transformation

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
pENTR1A Transformation not working CambridgeBiochemist Molecular Cloning Forum 1 06-18-2012 06:53 AM
Questions on rice transformation Wenxian Sun Arabidopsis and Plant Biology 0 04-21-2009 02:25 PM
PEG-mediated Arabidopsis Transformation Zhang, Yi Arabidopsis and Plant Biology 0 11-10-2006 11:05 PM
DNA for protoplast transformation Irina Teplova Arabidopsis and Plant Biology 0 08-16-2004 04:10 PM
high efficiency transformation Merijn Salverda Protocols and Methods Forum 1 12-23-2003 05:51 PM

All times are GMT. The time now is 04:27 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 0.11431 seconds with 16 queries