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Serum is the liquid component of blood that is obtained when cells and clotting factors are removed. It is widely used as a blocking agent for many different research applications as it contains antibodies and other proteins that can freely bind reactive sites in sample material. Serum is also popular as an antibody diluent, where it provides a more native environment for sensitive antibody reagents compared to using proteins such as bovine serum albumin (BSA) or fish skin gelatin. Another important application of serum is its use as a control for testing antibody purification methods.
How are sera used for immunoassay applications?
A typical immunoassay application such as Western blotting, immunocytochemistry (ICC), or immunohistochemistry (IHC) involves performing a sequential series of steps to enable target detection. Blocking is one of the earliest steps in the workflow and is usually followed by incubation with a primary antibody, washing to remove any unbound antibody reagent, and incubation with a labeled secondary antibody, prior to further washing and target visualization.
The blocking step is critical to prevent non-specific binding of secondary antibodies that can cause unwanted background signal, leading to false-positive results. It can be achieved by incubating samples in 1-5% (v/v) serum for a pre-determined length of time, at a set temperature, allowing endogenous serum proteins to saturate any reactive sites. Where serum is employed as a blocking reagent, it is common practice for it also to be used for diluting primary and secondary antibody reagents, rather than switching to another blocker such as BSA.
When selecting sera for research use, it is important to match the host species to that of the secondary antibody. Serum from the host species of the primary antibody would be detected by secondary antibody reagents, generating an unwanted non-specific readout. In situations where secondary antibodies from several different hosts will be combined for detection (e.g., multiplexed IHC), it may be necessary to mix sera from all the host species to ensure effective blocking.
Why choose sera from ICL?
With over 40 years’ experience in producing high-quality primary and secondary antibodies, ICL understands what it takes to generate reliable immunoassay results. We offer a comprehensive range of normal animal sera for both routine use and bulk purchase, all of which are sterile filtered and supplemented with sodium azide as a preservative. Like every ICL product, our sera are backed by rapid customer service and troubleshooting advice from our knowledgeable US-based team, helping you to generate results you can trust.