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Author Topic: Increasing DAB staining intensity  (Read 9403 times)

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Offline Payam

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Increasing DAB staining intensity
« on: January 05, 2008, 04:28:02 AM »
I used to work with liquid DAB chromogen from DAKO and always got perfect staining. Now that I have switched to DAB tablets, my staining has become very pale. Has anybody any comments on how to increase staining intensity when using DAB tablets? I am following the exact protocol suggested by company, one tablet dissolved in 10 ml tris-HCl buffer, aliquot and freeze in -20 and then thawing and using it in few days after making. I add 8 micro liter hydrogen peroxide (to 1 ml solution) and apply that to the slides for an incubation time of 30 min.
I have thought of reducing the amount of the buffer, i.e. adding 1 tablet (10 mg) to 5 ml buffer, instead of 10. Do you think this helps? is there any other suggestion?
regards,
Payam

Increasing DAB staining intensity
« on: January 05, 2008, 04:28:02 AM »

Offline richard03

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Re: Increasing DAB staining intensity
« Reply #1 on: January 06, 2008, 01:10:13 PM »
I would suggest to always use freshly prepared solution. Once the aliquot is taken out from freezer, you need to use it immediately.

Another suggestion is to double or triple the volume of the H2O2.

Offline ImmunoNYC

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Re: Increasing DAB staining intensity
« Reply #2 on: January 19, 2009, 04:16:33 PM »
I suggest you switch back to DAKO DAB, don't waste your time.

Re: Increasing DAB staining intensity
« Reply #2 on: January 19, 2009, 04:16:33 PM »