For both medicine and biology, immunology is an important field. Immunology is the study of diseases resulting from disorders of the immune system.
An invading organism, which causes a person or animal to be sick, is attacked by the immune system until they feel better.
Animal science researchers are continually searching for ways to boost animal immune systems through the development of high-quality products. To do this, researchers need to understand how certain factors affect the immune system. For example, genetic variation, growth and development, and other species-specific health factors, such as reproduction, digestion, and metabolism.
Similar to a human, animals can develop autoimmune diseases, cancer or many other diseases (for example, metabolic, cardiovascular, and neurodegenerative conditions) if their immune system is not functioning properly.
ELISA and Animal Immunology
Peptides, proteins, antibodies, and hormones can be detected and measured in the blood through a test called an enzyme-linked immunosorbent assay (ELISA).
ELISA is the most commonly used technique and works by coupling an antibody or antigen to an assay enzyme. Antigens and substances that can stimulate the production of antibodies are detected through the combination of the specificity of the antibody and the sensitivity of the assay. The plate is coated with high-affinity antibodies to enhance the sensitivity and precision of the assay.
ELISA can be used by animal researchers to assess whether an animal has antibodies related to particular infectious conditions. Antibodies are made in response to infection. As such, if antibodies are present in the bloodstream then this may indicate that an animal has been in contact with a particular virus.
The Indirect Method of ELISA
Antibodies can be tested for in the blood through several ELISA methods. The focus of this article is the most frequently used indirect method.
If the blood of a chicken needs to be tested then a fresh sample will be extracted and centrifuged. The supernatant fraction will be collected, then the sample will be diluted with a phosphate buffer solution at 1:100, and left to stand.
A solid-phase antigen is then formed by coating the antigens to create specific antigens, which link with solid-phase carriers. The sample is incubated overnight at 4 °C. The next day, 5% skim milk powder is added for sealing and is washed three times to remove unbound antigens and impurities.
The serum being tested (the first antibody) is added in the third step and incubated for one hour at 37 °C. Phosphate-buffered saline is then added and washed for 5 minutes at room temperature before drying up.
In step four, enzyme-labeled antibodies are added and incubated for one hour at 37 °C. Then phosphate-buffered saline is added and washed for 5 minutes at room temperature before drying up.
Finally, a substrate for the chromogenic assay is added and the reaction is kept at 37 °C for 30 minutes. The test is finished by adding 2 ml of sulfuric acid solution. The optical density value can now be measured using the ELISA detector and the ratio of positive serum to negative serum calculated.
Samples are required to be shaken and blended several times throughout the testing process with the ELISA method described here. To ensure the durability and uniformity of the experiment samples must be properly blended before and after dilution, and all reagents must be shaken before adding them in. This process can be carried out efficiently and effectively using the OHAUS Microplate Shaker.
It also proves a useful tool during the washing process, where antigens are separated or antibodies bound to the solid phase from non-specific components adsorbed during incubation. This part of the testing process is just as important to meet the specificity of ELISA.
The OHAUS Microplate Shaker
A valuable asset in any lab wishing to conduct ELISA testing, the OHAUS Microplate Shaker is compact in design, meaning that it can easily fit into hoods or incubators. Its rubber base ensures durability and provides added stability. Consistent uniform shaking action and reliable results are provided through the microprocessor control and brushless DC motor.
The speed-ramping feature of the OHAUS Microplate Shaker can increase the rate of speed slowly and safely to the desired set-point. Furthermore, when the system detects an obstruction or overload of the tray audible and visual signals activate.
Learn more about OHAUS shakers.
About OHAUS Corporation
OHAUS Corporation manufactures an extensive line of weighing products, laboratory equipment and analytical instruments that meet the weighing and measurement needs of virtually every industry.
They are a global leader in the laboratory, industrial and education channels, as well as a host of specialty markets, including the food preparation, pharmacy and jewelry industries. An ISO 9001:2008 manufacturer, OHAUS products are precise, reliable and affordable, and backed by industry-leading customer support.
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