Abcam Ad

Author Topic: BRDU  (Read 6392 times)

0 Members and 1 Guest are viewing this topic.

Offline Annica Andersson

  • SuperTech
  • ****
  • Posts: 21
BRDU
« on: September 19, 2007, 03:15:53 AM »
Hello,
We have perfused rats with 4%PF/picric acid and now we have problems to get the BrdU AB to work (Harlan, 1:100 for 90 min), 2M HCL 37C for 30 min might not be enough. When develope with DAB the staining looks weak and the ab dont penetrate to the nuklea. Its there a nother way to denature the sectons (12um) we also have big problems with DAB background and that does not īmake it easy to evaluate.
Regards Annica

BRDU
« on: September 19, 2007, 03:15:53 AM »

Offline richard03

  • Administrator
  • GoldMember
  • *****
  • Posts: 1212
Re: BRDU
« Reply #1 on: September 19, 2007, 07:52:15 AM »
Citrate buffer antigen retrieval is an alternate method for BrdU antibody

http://www.ihcworld.com/_protocols/epitope_retrieval/citrate_buffer.htm

Offline dutchmaninvienna

  • SuperTech
  • ****
  • Posts: 29
Re: BRDU
« Reply #2 on: September 21, 2007, 02:03:27 AM »
It is a bit unclear how you have processed the sections, are they cryostate or paraffin sections? If you feel that you have problems with penetration of the antibodies you might 1) increase the incubation times, 2) use detergents, 3) in case of paraffin, increase the deparaffination times. 4) in case of cryostate sections use acetone or chloroform to permeabilize the membranes in your tissues. etc etc. Just shortly write down you protocol and it will be easier to find out your problem.

jan
Jan Bauer, PhD
Associate Professor
Div. of Neuroimmunology
Center for Brain Research
Medical University of Vienna, Austria

Re: BRDU
« Reply #2 on: September 21, 2007, 02:03:27 AM »