This kit contains all of the reagents necessary to prepare either a DAB or a DAB/nickel substrate working solution. Vector DAB Substrate (3,3’-diaminobenzidine) produces a brown reaction product in the presence of peroxidase (HRP) enzyme. This kit also includes nickel chloride, which, when added to the substrate working solution, results in a gray-black reaction product. Vector DAB Substrate is suitable for darkfield and electron microscopy (EM).
Features of this kit:
Ideal for IHC/ICC/ISH and blots
Ideal for single and multiple labeling
No wait times for mixing and dissolving powders or tablets
Sufficient reagents to produce 300 ml of working solution
Immunohistochemistry / Immunocytochemistry, In situ hybridization, Blotting Applications, Elispot
I am performing IHC on tissue sections using a mouse IgG primary antibody. My detection system is a biotinylated anti-mouse IgG secondary antibody, a VECTASTAIN® Elite ABC peroxidase kit (PK-6100), in combination with a DAB substrate kit (SK-4100), and I do not see any staining. How do I know if the end detection reagents are working?
Make a fresh working solution of DAB substrate per instructions. Place a small volume (~1 mL) of this DAB substrate into a clean glass test tube. To this 1 ml aliquot, add one drop (~50 µL) of Reagent B only from the VECTASTAIN® Elite ABC kit. If the HRP enzyme is active and the DAB reacts with it, an immediate color change will be observed. This indicates the end detection reagents are working appropriately.
This kit contains stock solutions in convenient dropper bottles. Slides developed with DAB/ Ni should be dehydrated, cleared, and permanently mounted. For additional information, view this product review on Biocompare.com.
Counterstain / Substrate Compatibility
This table is designed as a reference to determine the optimal counterstain / substrate combination for your application. Consideration should be given to tissue type, antigen unmasking protocol, and other detection parameters to achieve the desired staining intensity.
*This substrate shows a slight decrease in sensitivity following the methyl green protocol. This decrease can be minimized by reducing the heat incubation and acetone rinse times. **Substrate dissolves in acetone wash. ***A slight color change in ImmPACT NovaRED and Vector NovaRED reaction produt may be seen using methyl green.