antibodies cross reacting with TdTomato

[Matthias Zehner]

Hello,

I have just a short question about labelling TdTomato with an antibody. I'm
trying to image an transgenic parasite which is expressing TdTomato and have
the problem, that I have to fix my probes with PFA to conserve the
tomato-fluorescence... unfortunately this impairs a lot of my other antibodies.

So I searched for antibodies that can be used to label TdTomato. I found
that some dsRed antibodies cross-react, but was not able to find out which
one do and which one can not recognise it.

Has somebody experience in this problem or can somebody recommend me an
antibody that might do this job?

Many thanks for your help!
Matthias Zehner


**************************************

Matthias Zehner, Dipl
Centre for Immunology and Infection
Department of Biology and Hull York Medical School
University of York

Email:  [hidden email]

[Michael Schell]

Hi Mathias,

We use the rabbit polyclonal against RFP sold by Rockland.  Recognizes tdTomato on PFA-fixed cells/tissues, and also on Westerns.  If you  don't dilute enough, you do see a non-specific nuclear stain.  Try  diluting to 1:2000  or more for specific labeling of tdTomato.

Mike



Michael J. Schell, Ph.D., CIV, USUHS
Assist. Professor
Dept. of Pharmacology
Uniformed Services University
4301 Jones Bridge Rd.
Bethesda, MD  20814-3220
tel:  [phone]
[hidden email]
>>> Matthias Zehner <[hidden email]> 05/22/09 9:01 AM >>>
Hello,

I have just a short question about labelling TdTomato with an antibody. I'm
trying to image an transgenic parasite which is expressing TdTomato and have
the problem, that I have to fix my probes with PFA to conserve the
tomato-fluorescence... unfortunately this impairs a lot of my other antibodies.

So I searched for antibodies that can be used to label TdTomato. I found
that some dsRed antibodies cross-react, but was not able to find out which
one do and which one can not recognise it.

Has somebody experience in this problem or can somebody recommend me an
antibody that might do this job?

Many thanks for your help!
Matthias Zehner


**************************************

Matthias Zehner, Dipl
Centre for Immunology and Infection
Department of Biology and Hull York Medical School
University of York

Email:  [hidden email]

[Matthias Zehner-2]

Many thanks for your quick answer! I will try this antibody out... do you know if its also
working on aceton-fixed tissues?

Again many thanks for your help
Matthias

[sgkcck]

It should

 

Thank you and we look forward to hearing from you

Sincerely

Stacie Kirsch
Electron Microscopy Sciences
1560 Industry Road
Hatfield Pa 19440
Tel: [phone] Fax: [phone]
E-Mail: [hidden email] or [hidden email]
Web: www.emsdiasum.com

 

In a message dated 5/25/2009 3:14:08 P.M. Eastern Daylight Time, mazehner@... writes:

Many thanks for your quick answer! I will try this antibody out... do you know if its also
working on aceton-fixed tissues?

Again many thanks for your help
Matthias

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