(no subject)

[Indig, Fred (NIH/NIA/IRP) [E]]

Dear Neeraj,

The fixation, not membrane permeabilization, seems to be more of a factor for nuclear proteins.

Try Methanol (100% formaldehyde-free at -20C for 20 min). Compare with 3.7% FA at RT.

Contact me off-list if you need further details.

Besrt Wishes,

Fred E. Indig, Ph.D.

Head, Confocal Imaging Facility

Hi List,

 

One of my colleagues is trying to visualize a nuclear protein using primary antibodies. Is there a special trick  or protocol that allows the primary and the fluorescent secondary antibodies better access to proteins inside the nucleus? They have already considered using either Triton or Tween-20.

 

Thanks,

 

Neeraj.

 

Neeraj V. Gohad, Ph.D.

Postdoctoral Fellow

Okeanos Research Group

Department of Biological Sciences

132 Long Hall

Clemson University

Clemson,SC-29634

Phone: [phone]

Fax: [phone]

 

Please note my new email address: [hidden email]

 

 

-- 

[Neeraj Gohad-2]

Dear Fred,

 

Thanks for the reply, I forwarded your suggestion to my colleague, and would much appreciate any further suggestions.

 

Best,

 

Neeraj.

 

Neeraj V. Gohad, Ph.D.

Postdoctoral Fellow

Okeanos Research Group

Department of Biological Sciences

132 Long Hall

Clemson University

Clemson,SC-29634

Phone: [phone]

Fax: [phone]

 

Please note my new email address: [hidden email]

 

 

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Fred E. Indig, Ph.D.
Sent: Thursday, January 28, 2010 1:30 PM
To: [hidden email]
Subject:

 

Dear Neeraj,

 

The fixation, not membrane permeabilization, seems to be more of a factor for nuclear proteins.

 

Try Methanol (100% formaldehyde-free at -20C for 20 min). Compare with 3.7% FA at RT.

 

Contact me off-list if you need further details.

 

Besrt Wishes,

 

 

Fred E. Indig, Ph.D.

Head, Confocal Imaging Facility

 

Hi List,

 

One of my colleagues is trying to visualize a nuclear protein using primary antibodies. Is there a special trick  or protocol that allows the primary and the fluorescent secondary antibodies better access to proteins inside the nucleus? They have already considered using either Triton or Tween-20.

 

Thanks,

 

Neeraj.

 

Neeraj V. Gohad, Ph.D.

Postdoctoral Fellow

Okeanos Research Group

Department of Biological Sciences

132 Long Hall

Clemson University

Clemson,SC-29634

Phone: [phone]

Fax: [phone]

 

Please note my new email address: [hidden email]

 

 

 

 

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