# DAB staining Immunohistochemistery

**URL:** <https://forum.painresearcher.net/t/dab-staining-immunohistochemistery/1353>\
**Category:** Microscopy (IHC/ISH/ICC/In Vivo)\
**Tags:** immunohistochemistry\
**Created:** [April 4, 2025, 7:48pm UTC](https://forum.painresearcher.net/t/dab-staining-immunohistochemistery/1353 "2025-04-04T19:48:57Z")\
**Posts on this page:** 3\
**Page:** 1

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**Author:** ![HebaReda](https://avatars.discourse-cdn.com/v4/letter/h/aca169/32.png) [@HebaReda](https://forum.painresearcher.net/u/HebaReda)\
**Post date:** [April 4, 2025, 7:48pm UTC](https://forum.painresearcher.net/t/dab-staining-immunohistochemistery/1353/1 "2025-04-04T19:48:57Z")

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Hi,  
I am planning to use DAB with nickel in immunohistochemistery so what is the best concentration that should be used please as I am confused with that?  
I was preparing it freshly by adding 2.7 mg in 10 ml PBS (so a concentration of 0.027 % w/v) and then nickel (0.05% w/v) and H2O2 (0.05%) but I did not get a good stain so should I increase DAB to 5-6 mg?  
I filter it with 0.45 micro filter before adding H2O2.  
I am working on mouse spinal cord sections.

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**Author:** ![achamess](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.painresearcher.net/achamess/32/534_2.png) [@achamess](https://forum.painresearcher.net/u/achamess)\
**Post date:** [April 6, 2025, 11:03am UTC](https://forum.painresearcher.net/t/dab-staining-immunohistochemistery/1353/2 "2025-04-06T11:03:33Z")

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i’ve had good results with this: [https://vectorlabs.com/products/dab-hrp-substrate/?gad\_source=1&gbraid=0AAAAAD\_Ur5gao0LUf2NxUr\_phHuzFxbPB&gclid=CjwKCAjwzMi\_BhACEiwAX4YZUM6QJY-PNqJ75dbfDh-3Pt\_zfInM9KW6nOv012I3ThzWnnHttRbM5xoCLsYQAvD\_BwE](https://vectorlabs.com/products/dab-hrp-substrate/?gad_source=1&gbraid=0AAAAAD_Ur5gao0LUf2NxUr_phHuzFxbPB&gclid=CjwKCAjwzMi_BhACEiwAX4YZUM6QJY-PNqJ75dbfDh-3Pt_zfInM9KW6nOv012I3ThzWnnHttRbM5xoCLsYQAvD_BwE)

See what guidance you might find from their docs.

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<div class="post-metadata">

**Author:** ![HebaReda](https://avatars.discourse-cdn.com/v4/letter/h/aca169/32.png) [@HebaReda](https://forum.painresearcher.net/u/HebaReda)\
**Post date:** [April 6, 2025, 9:20pm UTC](https://forum.painresearcher.net/t/dab-staining-immunohistochemistery/1353/3 "2025-04-06T21:20:39Z")

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Hi Alex,  
Thanks so much for your reply and advice. I will try this kit.