What is Western Blot Service?What is Western Blot Service?
Western blot service remains the foremost method for protein detection. It is a technique that separates proteins by molecular weight and then probes the blot with antibodies to analyze the presence or approximate amount of specific proteins. Unlike PCR (polymerase chain reaction) and RT-PCR (reverse transcription-PCR), which address gene level information, western blot analysis provides insights into post-transcriptional regulation, modifications and protein abundance.
Polyacrylamide gel western blot service (PAGE) is a common technique that separates charged biomolecules such as proteins by physical properties (particle size, charge and mass) by passing them through a gel under the force of an electric current. After separating the proteins, they are transferred to a PVDF membrane for identification and documentation.
Once the proteins are in the membrane, they can be reversibly stained with a protein stain such as Ponceau S stain to verify that they have been transferred properly. However, this method has limited sensitivity and often fades during storage or with time limiting its use for documentation.
More recently, fluorescence detection has been gaining popularity with researchers. This involves probing the blot with an antibody that has been conjugated to a fluorescent dye such as horseradish peroxidase (HRP) or a fluorophore, such as infrared or near-infrared spectral-shifted antibodies. This approach has the advantage of reducing background, but requires specialized equipment and expensive fluorophore-conjugated antibodies and is not as sensitive as chemiluminescence. Ichor offers a convenient and sensitive alternative with the Azure c600 imaging system which allows you to capture chemiluminescent and fluorescent images of your blots without the need for a dark room, timer or educated guesses about exposure times.
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