For instance, the ABHD10 polyclonal antibody is important for cellular processes such as nuclear signaling, immunochemistry, immunology, signal transduction, autophagy antibodies etc. To determine the presence or absence of these polyclonal antibodies, the following processes will be necessary:
Western Blot
This is a technique used in cellular and molecular biology. Proteins from a mixture can be easily identified from a mixture of complex proteins using the Western Blot. Three principles are applicable for success of the technique and they include separation by size, transfer of the extract on to a solid support and marking the target proteins using the most appropriate primary and secondary antibodies.
Separation of the proteins is done through gel electrophoresis and the extracted protein molecules are transferred to a membrane where bands will be produced. Each band represents a protein but to get the target protein, you should incubate the membrane with label antibodies, specific to target protein.
You will have to wash off any unbound antibodies and have bound antibodies detected by the developing film. The thicker the band on the film, the higher the amount of protein present.
EIA/ ELISA
Enzyme-linked immunosorbent assays, also referred to as enzyme immune assays are biochemical techniques for detection of antibodies or antigens in given samples. It is used widely as a diagnostic tool for plant pathology and medicine.
Generally, ELISA/ EIA tests involve an unknown amount of antigens affixed to a well-plate with 48 or 96 wells. Thereafter, a specific antibody, for example the polyclonal antibody, ABHD10 Antibody is applied over the surface for it to bind to the antigen. The antibody introduced into the antigen is enzyme-linked. Finally, a substrate is added to the mixture for signal detection. Normally, the antigen-antibody interaction is shown by colour change in the chemical substrate with the antigen-antibody complex.
Sandwich EIA is the most suitable for polyclonal antibodies and polystyrene is the best and most preferable substrate to use thought the thickness will be determined by the size of the plates.
Immunohistochemistry
This is a histological procedure used in detection of diseases ailing the cells, tissues or organs. It is mainly used to detect the presence or absence of a specific protein marker for diagnosis and classification of tumors. The detection process is dependent on the principles of the antibody that binds specifically to that particular antigen in tissues. IHC utilizes staining to identify target molecules and their distribution. Sample separation and labelling are the main activities involved in IHC tests for diagnosis. Polyclonal and monoclonal antibodies are used in detection.
There are two main methods of IHC detection i.e. the direct and the indirect methods. The direct involves direct reaction of the antibody with the antigen and only one antibody is used. The later utilizes two types of antibodies and is therefore more specific. You must use a stain to visualize the results.
In conclusion, use of polyclonal antibodies gives more rapid results because of the numerous epitomes available. This makes diagnosis of diseases like cancer, cardiovascular diseases, and neurological disorders fast and accurate.
- Christie Wilcox
- Publisher: Scientific American / Farrar, Straus and Giroux
- Edition no. 0 (08/15/2017)
Last update on 2020-01-27 / Affiliate links / Images from Amazon Product Advertising API
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