Thermo Scientific Pierce Metal-Enhanced DAB Substrate Kit uses cobalt and nickel chloride in a special formulation of diaminobenzidine peroxidase substrate that yields intense color for immunohistochemical staining.
- Incredible sensitivity – fifty times more sensitive than the traditional DAB method and thiry times more sensitive than other metal-intensified versions of DAB
- Low background, high intensity – get a crisp dark brown-black precipitate that is more intense than the dull brown precipitate when using DAB without enhancement. And, even with the increased intensity, background is almost nonexistent
- Only two components – simply mix the two liquid components and your working solution is ready to use
- Six-hour stability – the innovative working solution is stable for more than six hours at room temperature, while other DAB substrates must be used immediately
To prepare the product, the 10X Metal Enhanced Substrate Solution is diluted with the Pierce Stable Peroxide Buffer prior to use, and the special formulation allows the 1X substrate to be stable for at least 6 hours at room temperature. The background color of the substrate does not increase and, unlike typical metal enhanced DAB methods, the reactivity is unchanged (1,2). The resulting precipate is a dark brown-black that is more intense than the brown color from DAB alone. The intensity not only increases, but the background is also reduced, which increases the signal-to-noise ratio and makes screening tissue or membrane antigens easier.
The detection limits of various DAB substrates.
||Lower Detection Limit (5)
|DAB Substrate (3)
||0.800ng Horseradish peroxidase-conjugated streptavidin
|Metal Enhanced DAB Substrate (4)
||0.550ng Horseradish peroxidase-conjugated streptavidin
|Pierce Metal Enhanced DAB Substrate Kit
|0.017ng Horseradish peroxidase-conjugated streptavidin
|Prostatic acid phosphatase stained with Pierce Metal Enhanced DAB and blocked with SuperBlock Blocking Buffer in TBS.
||Prostatic acid phosphatase stained with DAB according to the method by Graham and Karnovsky, 1966 (4).
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- Adams, J.C. (1981). J. Histochem. Cytochem. 29: 775.
- Georghegan, W.D. and Ackerman, G.A. (1977). J. Histochem. Cytochem. 25: 1187.
- Graham, R.C. and Karnovsky, M.J. (1966). J. Histochem. Cytochem. 14: 291-302.
- Heimberger, A.B., et al. (2003). Clin. Cancer Res. 9: 4247-4254.
- Rodbard, D. (1978). Anal. Biochem. 90: 1-12.
- Wanaget, J., et al. (2001). FASEB J. 15: 322-32.
Immunohistochemistry (IHC) technical guide
IHC immunodetection strategies technical guide
Other Immunohistochemistry Reagents
Formaldehyde (16% solution)