I want to do a double staining on some IHC samples, can I use 2 different Peroxidase detection systems as AEC and DAB together? Which mounting media should I use (if the staining is possible)?
Hi Almudena
Yes, You can use two different peroxidase chromogens that is AEC and DAB. In such case note that AEC (red) and DAB (brown) are compatible and can be used simultaneously in dual staining procedure. In some cases AEC show incompatibility with DAB (when it produce Gray-Black color due to nickle enhancement ) due to various reasons. And you can use aqueous mounting in this.
Hope it helps
Thanks Nasir, As I have never done it, I should first do the staining for one antibody/chromogen and after the next right?
Yes you are right! Optimize conditions for one antibody first. You can start different conc of your pri-ab according to data sheet provided. Let say start with different conc like 1/50, 1/100, 1/150 and 1/200 on positive control first (dont forget to use negative controls, use PBS in place of Pri-ab). You will get different degrees of staining from light to dark colors, choose the most appropriate stain intensity for your purpose and note the conc of Ab used. Repeat the same procedure for 2nd Ab and note conc of both Abs, then combine these conc in dual staining procedure. Good luck
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining