Product Name
Diaminobenzidine Cobalt Chromogen Kit
Catalog Number
GTX73331
Product Description:
Diaminobenzidine ( DAB ) Cobalt Chromogen Kit
Background:
The use of 3-3' diaminobenzidine tetrahydrochloride (DAB) as a chromogen, when performing peroxidase-based immunocytochemical procedures, provides the means of organic solvent-stable permanent chromogen depositions. Therefore, specimens developed with DAB can be dehydrated and mounted with organic solvents-based plastic polymers such as Permount or Clarion (Cat. No. GTX73339 and GTX73340). Since DAB is a suspected carcinogen, many laboratories avoid the handling of the powdered chemical. However, the availability of DAB tablets minimizes the risk of exposure and facilitates its use in the immunohistology laboratory. Our Diaminobenzidine Cobalt Chromogen is significantly more sensitive than the regular DAB.Cobalt ions included in the concentrated buffer formulation change the regular DAB color. The DAB Cobalt depositions are black in color and can be counterstained with alcohol-based hematoxylin or with Our Aqueous Hematoxylin (Cat. No. GTX73341). Nuclear Fast Red or Methylene Green can be also used as counterstains.**When using Nuclear Fast Red or Methylene Green, dehydrating the tissue sections and mounting with Clarion or another organic based mounting medium, is required. Aqueous mounting media like Crystal/Mount are not compatible with these counterstains. These products are designed to produce a color deposition on tissue sections processed by immunocytochemical techniques using alkaline phosphatase or peroxidase as tracers. When using these products, it is very important to select the appropriate chromogen substrate for the enzyme tracer being used in the assay. One mL of final working solution will yield approximately 5-6 tissue sections. The number of tests performed will vary depending on the application. Estimations are derived from use with the capillary-action based MicroProbe(tm) system, which draws 150-200 mL per test. Each 20-tablet kit contains sufficient reagents to make 100 mL of final working solution. The 100 tablet chromogen-only kit will yield 500 mL of final working solution. All other kits contain sufficient reagents to make 300 mL of final working solution. Working Chromogen Preparation: Remove the DAB Cobalt Chromogen kit from the refrigerator 20 minutes before the preparation of the working chromogen solution. This will minimize the condensation of ambient water on the tablets during handling. 1.Transfer 5 mL of distilled water into suppliedplastic bottle. 2.Add 5 drops of Concentrated Buffer Reagent.Mix. 3.Add one chromogen tablet. Allow tablet to dissolvefor approximately 5 minutes with occasional mixing.Secure the cap of the bottle containing the unusedtablets and return it to the original plastic box. 4. After tablet dissolves, add one drop of ConcentratedSubstrate. Mix well. 5.Apply this solution to the specimens. Make sureto wash the specimens extensively with buffer toremove tracing reagent containing peroxidase. 6.Incubate for 5-10 minutes at room temperature.Wash specimens with distilled water and proceedwith the counterstaining steps.
Storage Instruction:
Store at 4 °C. Do not freeze.
Tested Applications:
Immunohistochemistry. The usefulness of this product in other applications has not been determined.
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