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).
Unit Size | 1 kit |
---|---|
Applications | Immunohistochemistry / Immunocytochemistry, In situ hybridization, Blotting Applications, Elispot |
Detection Enzyme(s) | Peroxidase |
Substrate Reaction Product | Precipitating Reaction Product |
Chromogen Color | Brown, Gray-Black |
Microscopy | Brightfield, Darkfield, Electron, Spectral Imaging |
Additional Substrate Properties | Heat Resistant, Multiple Labeling |
Mounting | Aqueous (Hardening), Non-Aqueous (Permanent) |
Safety Title | WARNING: Cancer – www.P65Warnings.ca.gov |
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.
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.
Substrate | Catalog Number | Hematoxylin (H-3404) Hematoxylin QS (H-3401) | Methyl Green (H-3402) | Nuclear Fast Red (H-3404) |
ImmPACT DAB (brown) | SK-4105 | Excellent Contrast | Excellent Contrast | Fair Contrast |
ImmPACT DAB EqV (brown) | SK-4103 | Excellent Contrast | Excellent Contrast | Fair Contrast |
DAB (brown) | SK-4100 | Excellent Contrast | Excellent Contrast | Fair Contrast |
DAB-Ni (gray-black) | SK-4100 | Excellent Contrast | Fair Contrast* | Good Contrast |
ImmPACT AEC (red) | SK-4205 | Excellent Contrast | Counterstain Incompatibility** | Color Incompatibility |
ImmPACT AMEC Red (red) | SK-4285 | Excellent Contrast | Counterstain Incompatibility** | Color Incompatibility |
AEC (red) | SK-4200 | Excellent Contrast | Counterstain Incompatibility** | Color Incompatibility |
TMB (blue) | SK-4400 | Color Incompatibility | Counterstain Incompatibility | Excellent Contrast |
ImmPACT VIP (purple) | SK-4605 | Fair Contrast | Excellent Contrast | Poor Contrast |
Vector VIP (purple) | SK-4600 | Fair Contrast | Excellent Contrast | Poor Contrast |
ImmPACT SG (blue-gray) | SK-4705 | Poor Contrast | Good Contrast | Excellent Contrast |
SG (blue-gray) | SK-4700 | Poor Contrast | Good Contrast | Excellent Contrast |
ImmPACT NovaRED (red) | SK-4805 | Excellent Contrast | Excellent Contrast*** | Color Incompatibility |
Vector NovaRED (red) | SK-4800 | Excellent Contrast | Excellent Contrast*** | Color Incompatibility |
ImmPACT Vector Red (magenta) | SK-5105 | Excellent Contrast | Excellent Contrast | Color Incompatibility |
Vector Red (magenta) | SK-5100 | Excellent Contrast | Excellent Contrast | Color Incompatibility |
Vector Black (black) | SK-5200 | Excellent Contrast | Excellent Contrast* | Excellent Contrast |
Vector Blue (blue) | SK-5300 | Color Incompatibility | Good Contrast | Excellent Contrast |
BCIP / NBT (indigo) | SK-5400 | Color Incompatibility | Excellent Contrast* | Excellent Contrast |
*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. |
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