PEROXIDASE SUBSTRATES (HRP SUBSTRATES)

In general, HRP-substrates produce sharper, denser precipitation products with crisp localization. Some substrates are versatile and allow non-aqueous as well as aqueous mounting (DAB, SG). Other substrates are less versatile and will work on either permanent or aqueous mounts, but not both. HRP substrates also vary in their ability to provide contrast with common counterstains. Finally, for multiplexing (multiple antigen labelling) experiments, a proper combination of substrates should be carefully chosen, in order to provide best contrast results.

DAB (3,3´-diaminobenzidine) is probably the most commonly used HRP-substrate. DAB is popular due to its robust and sensitive staining, high stability and excellent contrast with hematoxylin and methyl green counterstains.

Picture: Human tonsil (FFPE serial sections) stained for CD20 (1:200), detected with ImmPRESS HRP anti-mouse IgG. The only variable reagent was DAB substrate.

 

Our HRP substrates:

• Brown: Vector DAB, ImmPACT™ DAB, ImmPACT™ DAB EqV
• Gray-Black: Vector DAB + Ni
• Purple: Vector VIP, ImmPACT™ VIP
• Blue-Gray: Vector SG, ImmPACT™ SG
• Red: Vector NovaRED, ImmPACT™ Novared, Vector AEC, ImmPACT™ AEC, ImmPACT™ AMEC Red
• Blue: TMB

Table 1