Immunoblotting (alternatively, Western blot, line-ELISA, dot-blot or cold-blot) is an analytical technique used to detect specific proteins in a patients’ sample. The proteins to be analyzed are dispensed onto a membrane (typically nitrocellulose, nylon or PVDF), where they are probed by the antibodies of the patient. A simple secondary antibody conjugated to a reporter dye forms the basis of the color reaction which can be read by eye (qualitative) or instrument (qualitative and quantitative).
In the field of diagnostics their use is widespread as a screening tool where a patient can be profiled against multiple markers simultaneously. Common Immunoblot screening applications include:
- Cancer Biomarkers
- Autoimmune diseases
- Food intolerance
- Sexually Transmitted Diseases (STDs)
BioDot is at the forefront of the production of Immunoblot diagnostic tests, from small batch R&D platforms up to large multi-dispenser (32 channels) batch platforms or Reel to Reel systems for full scale production. Typically proteins (and sometimes DNA) are deposited by a contact or non-contact BioJet Plus dispenser. Once dispensed the ‘blots’ are blocked then dried. The product is then cut into small strips for their assembly into cassettes.
- FrontLine or BioJet technology for dispensing the lines
- AirJet equipped dispensing platforms for blocking
- Forced air ovens (with IR sensors) for drying
- Guillotine, rotary cutters and sheet slitters for singulation of tests.
Additional Information ...
Using electrophoresis gel in the analysis of specific antibodies with sampled proteins.
Immunoblot testing is a procedure which involves the use of electrophoresis gel to separate proteins which are then transferred to a nitrocellulose membrane where they can be observed or detected by the implementation of specific antibodies that react with the sampled protein. The target proteins usually originate from tissue homogenate or tissue extract that, when applied to the nitrocellulose membrane, separate into either a 3d protein structure which indicates a native protein or a polypeptide structure in which its length is measured to indicate a denatured protein.
Immunoblot testing is used in various applications which include the detection of antibodies in human serum to indicate the presence of a disease or virus, such as the Human immunodeficiency Virus or HIV, Lyme disease, Hepatitis B, and many other diseases. Apart from useful application in the human specie, immunoblot testing can also be employed within several animal species as well.