Short communication

, Volume: 11( 9)

Antibodies as Probes for Proteins

*Correspondence:

Thomas Department of Analytical Chemistry, University of Cologne ,Germany, Tel: +93314852471; E-Mail: thomasuyd@gmail.com

Abstract

Introduction

Genetic and functional genetic studies require the discovery of not only DNA and RNA, but also certain proteins. In these studies, antibodies replace nucleic acid probes as reagents that can react selectively with different protein molecules. Immune proteins produced by antibodies (lymphocytes B) react against molecules (antigens) that an organism recognizes as foreign substances for example, the coat of viruses. The antibodies of vertebrates are able to produce millions of different antibodies, each of which is particularly sensitive to a different antigen, which may be protein, carbon dioxide, or a non-biological molecule. Each lymphocyte produces only one type of antibody, but different lymphocyte genes vary due to genetic rearrangement during immune development. These mutations trigger a series of lymphocytes that have different immune systems, which are programmed to respond to different antigens.

Antibodies can be produced by injecting an animal with any foreign protein. For example, antibodies against human proteins are often raised by rabbits. The sera of such vaccinated animals contains a mixture of antibodies (produced by different lymphocytes) that respond to multiple sites on the vaccine antigen. However, a single type of antibody (monoclonal antibodies) can also be produced by enlargement of clonal lines of lymphocytes B in vaccinated animals (usually mice). Because each lymphocyte is programmed to produce only one antibody, the clonal line of lymphocyte produces a monoclonal antibody that sees only one antigenic separator, thus providing a highly specific immunological reagent.Although antibodies may be elevated against proteins purified in cells, other active ingredients may be used for vaccination. For example, animals can be vaccinated with complete cells to boost the immune system against unknown proteins produced by a particular type of cell (e.g., cancer cell). Such antibodies may be used to identify proteins that are specific to the type of cells used for vaccination. In addition, antibodies are often elevated against proteins that are expressed in bacteria as clone regenerated. In this way, cellular cloning allows the production of antibodies against proteins that can be difficult to separate from eukaryotic cells. In addition, antibodies can be increased against synthetic peptides that contain only 10 to 15 amino acids, rather than resistant to whole proteins. Therefore, once the genetic sequence is known, antibodies to peptides are grouped together in part of their predicted sequence of proteins that can be produced. Because antibodies against these synthetic peptides are normally responsive to a stable protein, it is possible to produce antibodies against a protein that starts only in the sequence of the combined genes.

Antibodies can be used in a variety of ways to get protein in cell extracts. Two common methods are immunoblotting (also called Western blotting) and immunoprecipitation. Western blotting is another version of Southern blotting. Proteins in cell extracts begin to break down by size through gel electrophoresis.Because proteins have different shapes and charges, however, this process requires a modification of the mechanisms used for nucleic acid electrophoresis.

Proteins are separated by a process known as SDS-polyacrylamide gel electrophoresis (SDS-PAGE), which dissolves in a solution containing a detergent containing sodium dodecyl sulfate (SDS). Each protein binds many purifying molecules, converting the protein and giving the protein a whole new amount. Under these conditions, all proteins move to a fine electrode their migration levels are determined (like those of nucleic acids) in size only. Following electrophoresis, the protein is transferred to a filter, which is then allowed to react with antibodies against the interested protein. The antibody bound to the filter can be detected in a variety of ways, thus identifying the protein targeting.

Acknowledgemnt

None

Conflict of Interest

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