The Western Blot is an assay intended to detect a single protein in a mixture of proteins from cells, tissues, blood serum or saliva sputum. It requires an antibody specific to the protein that is intended to be detected.
Anewex support life science researchers with custom Western Blot protein analysis services in ease-of-use, speed and sensitivity as well as a better ability results. Our expert team can help with experimental design and implementation at any point in your project.
Anewex Experimental plan of Western Blot services:
– Total protein extraction from cells, tissues.
– Construction of Western Blots, immunostaining blots with any antibodies
- Proteins are separated by polyacrylamide gel electrophoresis, usually SDS-PAGE.
- The proteins are transferred to a sheet of blotting membrane (usually nitrocellulose, though other types of membranes such as PVDF can be used). The proteins retain the same pattern of separation they had on the electrophoresis gel.
- The blot is incubated with a generic blocking protein (such as milk proteins) to shield any remaining protein-binding sites on the membrane. An antibody is then added to the solution, which is able to bind to the target protein. The antibody has been conjugated to an enzyme (e.g. Alkaline Phosphatase or Horseradish Peroxidase) or sometimes a fluorescent dye or particle.
- The location of the antibody is revealed by incubating it with a colorless substrate that the enzyme-conjugate converts into a colored product, which can be observed by eye and imaged.
– Quantitative analysis of immunostaining
- Tissue or cells
- Purified protein (>50 microgram/sample, concentration > 1mg/ml) OR lysate samples (total protein >500 microgram/sample, concentration >1mg/ml OR collected cells / tissues / blood serum / saliva sputum.
- Primary antibody to detect target protein
Quantitative analysis is performed using our Western Blot quantification analysis software and high quality digital images can be delivered via email.
* Total protein extraction and quantification: 2 days
* Prepare the blotting sample and conduct Western blot: 3 day
* Quantification analysis: 1 day
* Making report: 3 days
* Total: 1-2 weeks