Some Applications Of Antibody Labeling Some Applications Of Antibody Labeling February 27
Nick Martin Jersey , 2014 | Author: Marcie Goodman | Posted in Education
Antibody labeling has been a great step in increasing the accuracy in chemical reactions. The process has made it possible to set up procedures that were previously considered impossible. This is because many protein substrates can be easily identified and isolated unlike in the past. The labels that are commonly used have the ability to fluoresce and are known as fluorophores.
There are two types of labelling known; in vitro and in vivo. The former occurs outside the body. It involves a chemical reaction between one amino acid and another that acts as a label. A chemical reaction forms a covalent bond between the two amino acids. For this reaction to take place there are several things that are needed: a polymerase, a labelled amino acid or nucleotide and an ATP molecule.
The other form of labelling is known as in vivo or metabolic conjugation. This type takes place in the body. Typically nucleic acids or amino acids are cultured for a couple of weeks to allow for the label to get attached to the proteins. As the labelling process takes place, RNA and DNA molecules undergo replication. Once the proteins have been identified, they are isolated, purified and used for other reactions.
One of the greatest causes for concern is the fact that the conjugation process may result in interference of the intrinsic activity of the antibody or protein. Depending on the type of label that is used the avidity may be affected to different extents or may even remain unchanged. There are a number of methods that can be employed to determine the residual activity after the combination between the protein and the label.
It is important to ensure that the ration between the labels and the substrate is maintained within the recommended value. An optimal combination ration ensures that all the substrates are labelled as the conjugation reaction takes place. When the number of labels for each molecule is too high, the fluorescence may not take place and this may pose a great challenge to the person performing the chemical reaction.
The use of this procedure in active site probes is one of the commonest applications. The probes are designed to bind to an active site found on an enzyme. They are attached to a detectable tag, a reactive group and a spacer arm. The probes are electrophilic and easily react with nucleophilic residues on enzymes to form covalent links. The probes are for these reasons, used to identify different types of enzymes.
There are many types of enzymes that make use of active site probes as labels including kinases, phosphatases, serine hydrolases, metalloproteases, and cytochrome enzymes and so on. Among the enzymes, the probes are very useful in assessing the degree of inhibition by various types of molecules. They also are used in assessing the intrinsic activity of individual enzymes. The enzymes may also act as labels for other types of proteins. Examples of proteins that act in this manner include horseradish peroxidase
Andre Johnson Texans Jersey , glucose oxidase and alkaline peroxidase.
Antibody labeling has many other applications and the list continues to grow by the day. There are ongoing efforts aimed at streamlining the procedure and improving the efficiency. At the same time, many manufacturers of reagents have incorporated the labels in their products.
There comes a time when people need extra help to find out about antibody labeling. We urge you to spend a minute exploring this informative and scientific web page at http:www.columbiabiosciences now.
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