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DAB Substrate Kit, Peroxidase (HRP), with Nickel, (3,3'-diaminobenzidine)

SK-4100
SKU Unit Size Price Qty
SK-4100 1 kit
$193.00
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Description

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
  • Heat Stable
  • Permanent mounting
  • 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

Specifications

More Information
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

Kit Components

What's in the box:

  • 6 ml DAB Reagent 1
  • 6 ml DAB Reagent 2
  • 6 ml DAB Reagent 3
  • 6 ml DAB Reagent 4 (Nickel Optional)

Documents

Product FAQs

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.

Related Products

Endogenous alkaline phosphatase (AP) and peroxidase (HRP) activities in frozen intestine revealed with Vector Red AP Substrate (red) and ImmPACT DAB HRP Substrate (brown) (left). Same substrates used on BLOXALL Solution-treated tissue (right). BLOXALL B BLOXALL® Endogenous Blocking Solution, Peroxidase and Alkaline Phosphatase Catalog #: SP-6000-100
Small Bowel: Cytokeratin 8/18 (m), ImmPRESS Anti-Mouse Ig Kit, DAB Substrate Kit (brown), Hematoxylin QS (blue). Hematoxylin QS Counterstain Catalog #: H-3404-100
Tonsil: Cytokeratin (m), ImmPRESS Anti-Mouse Ig Kit, ImmPACT DAB (brown) substrate. Hematoxylin (blue) counterstain. Hematoxylin Counterstain Catalog #: H-3401-500
Tonsil section showing specific cell staining (purple, Vector VIP) with Vector Methyl Green (green) counterstain. Methyl Green Counterstain Catalog #: H-3402-500
Tumor tissue section showing specific cytoplasmic cell staining (blue/gray, Vector SG) with Vector Nuclear Fast Red (red) counterstain. Nuclear Fast Red Counterstain Catalog #: H-3403-500

Citations

Technical Information

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.

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.

Relative sensitivity of substrates in IHC: Vector Laboratories

Home Horseradish Peroxidase (HRP) Substrates DAB Substrate Kit, Peroxidase (HRP), with Nickel, (3,3'-diaminobenzidine)