Detection of Horseradish Peroxidase (HRP): Western BLoT Chemiluminescent Substrate Series
The Western BLoT HRP Substrate series are chemiluminescent substrates for detecting horseradish peroxidase (HRP)-labeled antibodies used during Western blotting. All of the substrates contain two-components, a peroxide reagent and a luminol/enhancer solution. To use, equal volumes of the two solutions are mixed, the blot is incubated with the solution, and chemiluminescence is detected using film or digital imaging equipment. Takara Bio offers four types of HRP substrates for Western blot detection; please refer to the table in the At-A-Glance section to compare the substrates' specifications.
The Western BLoT Chemiluminescence HRP Substrate (Cat. # T7101A) provides sensitivity suitable for detection using X-ray film. Although it is the most basic substrate in Western BLoT HRP Substrate series, it produces a strong signal and provides high sensitivity with short exposure times.
The Western BLoT Quant HRP Substrate (Cat. # T7102A) is intended for Western blot detection using a CCD camera. The signal from Western BLoT Quant is linear with respect to protein amount over a broad range of concentrations. In addition, this substrate produces a strong, long-lived signal, allowing long exposure times to enable the detection of low-abundance proteins. Western BLoT Quant can also be used for X-ray film detection.
The Western BLoT Hyper HRP Substrate (Cat. # T7103A) is more sensitive than Western BLoT Quant and is suitable for quantitative detection of low-abundance proteins. Western BLoT Hyper is also appropriate in situations when the amount of primary antibody is limited and a high dilution factor is desired, or when the primary antibody has a relatively low binding affinity. Western BLoT Hyper can also be used for X-ray film detection.
The Western BLoT Ultra Sensitive HRP Substrate (Cat. # T7104A) is the most sensitive substrate in the series and is suitable for quantitative detection of very low-abundance proteins. It is possible to detect protein levels in the femtogram range using this product.