Abcam Ad

Author Topic: Wash out of DAB stain??  (Read 3721 times)

0 Members and 1 Guest are viewing this topic.

Offline amurr

  • Newbie
  • *
  • Posts: 2
Wash out of DAB stain??
« on: March 04, 2009, 10:40:56 AM »
Hi,
Our lab has been double IHC staining on frozen rat brain sections for cfos then GnrH for several years.
Recently we have encountered a loss of stain problem.  Our staining w/ Vector DAB and DAB/ Ni
works well then by the end of the week the stains appear to be washed out.  We have compared
our staining trays which are washed and re used to 'virgin' chamber slide to rule out any chlorine bleaching
effects, no difference, we've order fresh DAB, 2Ab, etc.  We most recently have increased our primary
dilution on the cfos and got better results but after washing the stain is less intense for a single stain
and almost non detecable in the double stain.  We have used the SAME protocol for years, and have published
on this protocol in the past. Now we are stumped... Any help??  Thanks.

Wash out of DAB stain??
« on: March 04, 2009, 10:40:56 AM »

Offline Cardio

  • Global Moderator
  • GoldMember
  • *****
  • Posts: 249
Re: Wash out of DAB stain??
« Reply #1 on: March 04, 2009, 04:29:59 PM »
Others on the forum have had similar problems with DAB dissapearing.

What mounting solution are you using and have you considered postfixing after the staining?

Offline amurr

  • Newbie
  • *
  • Posts: 2
Re: Wash out of DAB stain??
« Reply #2 on: March 09, 2009, 11:11:20 AM »
Thanks! We are using Permount from Fisher to mount with.  i don't know if we can post fix after staining
or not because we are staining for GnRH immed. following our cfos one, same tissues.  we recently found our
county did a chlorine flush on the water system so we are double checking ph and trying DAB fresh from Sigma
instead of Vector kit this week. 

Offline ImmunoNYC

  • Global Moderator
  • GoldMember
  • *****
  • Posts: 684
Re: Wash out of DAB stain??
« Reply #3 on: March 11, 2009, 10:41:02 AM »
I would call the company. Also my favorite DAB is the world is DAB+ from DAKO.

Offline CanuckPhD

  • Global Moderator
  • GoldMember
  • *****
  • Posts: 296
Re: Wash out of DAB stain??
« Reply #4 on: March 12, 2009, 03:03:56 AM »
If you determine the reason for your DAB disappearing please post it. I have never encountered this issue and am curious about it. I don't have any idea why this would occur as it has always been very stable in my lab.

Re: Wash out of DAB stain??
« Reply #4 on: March 12, 2009, 03:03:56 AM »