Scientist Solutions: International Life Science Community By Scientists For Scientists
Home » Forums » Cytology, Histology and IHC » Immunohistochemistry/ Immunocytochemistry » How to Block Endogenous Peroxidase for DAB staining ?

Thanks to our sponsors who make this site possible

How to Block Endogenous Peroxidase for DAB staining ?

RSS Feed

Would you like to save this topic, event, protocol or job so you can find it again easily?

Just click the "Save to My Lab Drawer" link and the item will be saved in the My Lab Drawer section of your bench space.

Available to members only. Please log in or register for your free account now.

Trinidad and Tobago

Send PM
See Mini bio

Status: Frog Egg
Frog Egg
Topic Started by Kisspep
on 6/17/2011 14:11 PM   
Reply to this post Go to the top of the page

I recently attempted my first DAB staining in the mouse brain. I used 40um thick sections from frozen brains and we used a "free floating" method. Some protocols advised to use 0.3% H2O2/PBS to quench endogenous peroxidase and others suggested 0.3% H2O2/100% methanol. So I used both. What I noticed is that those that were treated with the methanol solution appeared very "bubbly" whereas those in the PBS treatment were clear and neurons and structures were clearly visible.
Was my experience with the methanol due to some technical error on my part or is this a normal occurance for this tissue and I should use PBS instead ?


Send PM
See Mini bio

Status: Scientist Solutions Evangelist
Frog Laureate
Posted By pundir
on 11/15/2011 15:22 PM   
Reply to this post Go to the top of the page

 methonal may be harmful to some antigens for antigens sensitive to methanol and frozen sections use PBS containing  0.3-3% H2O2 it works best for quenching the endogenous peroxidase.

impossible! let's do it................


Send PM
See Mini bio

Status: Cancer, Diagnostics & Proteomics Moderator
Frog Laureate
on 11/15/2011 4:30 AM   
Reply to this post Go to the top of the page

Hi Kisspep

Methanol induces precipitation of proteins that may may have lead to the bubble like appearance. Im not sure but it May not be recommended for use with brain sections

Gaganjot Singh Truth seems so closer now......

As a Scientist Solutions member, you are able to register a positive vote for any topic which you believe is useful and relevant to our board or any reply which you believe is especially well worded and helpful.

By participating in the voting, you will be helping to identify the best topics & replies on the board.

You may vote once for any one post, and you may not vote for your own posts.

A post (topic or reply) will earn one "thumbs up" icon for every 10 votes received (up to 3 thumbs up), and the person who made the post will also earn two bonus points.

learn more about member points.