In immunoblotting assays a soluble substrate is converted to an insoluble or precipitating end product and deposited on a membrane at the site of the enzyme reaction. Precipitating, colorimetric substrates produce a signal that is visualized by the unaided eye. These images can also be recorded photographically or with an imaging system. KPL offers a line of precipitating substrates for colorimetric peroxidase and phosphatase detection. The choice of substrate depends on a number of factors, including the demands of the assay, the method of detection and the preferences of the researcher.
Colorimetric Blotting Substrates for Peroxidase Detection
TMB Membrane Peroxidase Substrate is the most sensitive colorimetric substrate for use in membrane assays. 4 CN Membrane Peroxidase Substrate is particularly suited for assays where background may present a problem. DAB Reagent Set and TrueBlue perform well on Western and dot blots and are also designed for immunohistochemical applications.
Colorimetric Blotting Substrates for Phosphatase Detection
|
Figure on right: A two-fold dilution series from 250 ng to 3.9 ng of mouse IgG was loaded onto a gel, then blotted onto nitrocellulose. KPL's substrates for alkaline phosphatase detection demonstrate low nanogram detection and low background in Western blotting. |
 |
|
FirePhos Membrane BCIP/NBT (1-comp)
|
KPL’s FirePhos and BCIP/NBT Substrates are effective colorimetric substrates for blotting detection of phosphatase conjugates. FirePhosTM produces a red precipitate and BCIP/NBT a purple reaction product for a choice of colors. Both substrates provide sensitivity in the low nanogram range with minimal background (see figure above).
Please see KPL's Overview of Colorimetric Blotting Substrates for assistance in comparing and choosing the best substrate for your application. | 
shopping cart
