In the Western blotting protocol, reagents such as blocking buffers are essential to getting clear and specific results as they prevent antibodies from binding to the blotting membrane nonspecifically. Rockland offers a variety of products made to be paired with western blot protocols in a variety of research applications.
Blocking Buffers
Made to prevent non-specific binding of antibodies to the blotting membrane that can lead to high background
Chemiluminescent Reagents
Use for horseradish peroxidase (HRP) based detection with a variety of protein targets while allowing for optimal signal-to-background ratio
Chromogenic Substrates
Use for sensitive colorimetric detection with a variety of protein targets
Fluorescent Reagents
Use for fluorescent detection with fluorochrome labeled antibodies to decrease background fluorescence and optimize signal
What is Western Blotting?
Western blotting is a widely used method for detection of a specific protein in a complex matrix, such as cell or tissue lysate (i.e. protein extracts). The Western blot assay uses gel electrophoresis (SDS-PAGE or native PAGE) to separate proteins according to molecular weight.
Western blot allows the transfer of proteins separated on a gel (SDS-PAGE) to a sticky membrane. Protein transfer is facilitated by electric current, negatively charged proteins and supporting reagents such as filter paper, transfer buffer, and a nitrocellulose or PVDF membrane. Detection of the target protein in a Western is carried out by antibodies. Various detection methods such as chemiluminescent or fluorescent detection are available.
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