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Western blotting is a widely used technique for the detection and analysis of proteins. It is frequently used to study single proteins but the advent of multiplexed, fluorescence-based detection enables the simultaneous detection of several proteins.
Western blotting, or immunoblotting, is used for both qualitative and quantitative analysis of protein expression in a wide variety of biological samples.
The Amersham Western blotting portfolio provides optimal solutions for routine protein identification with chemiluminescence as well as for accurate and reproducible quantitative WB with fluorescence detection.
In Western blotting, native or denatured proteins are first separated by gel electrophoresis according to polypeptide length (under denaturing conditions) or by the three-dimensional structure of the protein (under native/non-denaturing conditions).
The separated proteins are then transferred to a membrane, typically made of nitrocellulose or polyvinylidene difluoride (PVDF), where they are probed using antibodies specific to the target protein.
The target protein is then detected and visualized using an imaging system.
Ensure that your sample is adequately pure and in the best possible condition.
Separate proteins on the basis of size and/or charge.
After separation, transfer the proteins from the gel to a solid support membrane.
Use specific antibodies to target your protein of interest.
Choose the most suitable detection method and imaging solution for your needs.
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