page_title: Antibody
section_title: Research applications
hierarchical_section_title: Antibody / Research applications
caption_attribution_description: English: a This image is made from a Molecular Probes demo slide: Cells: bovine pulmonary arthery endothelial cells Blue: nucleus stained with DAPI Green: Tubulin (microtubles) stained with antibody Bodipy FL goat anti-mouse IgG (Indirect fluorescent antibody stain) Red: F-Actin stained with Texas Red X-Phalloidin Deutsch: Endothelzellen aus der Inneren Wand (Endothel) von Lungenarterien des Rindes unter dem Mikroskop. Die Zellkerne sind mit DAPI blau markiert. Die Mikrotubuli wurden über einen Antikörper grün markiert. Mit rot fluoreszierendem Phalloidin wurden die Aktinfilamente markiert. Français&#160;: Cellulles endothéliales vues au microscope. En bleu, noyaux marqués au DAPI. En vert, microtubules marqués par un anticorps. En rouge, actine marquée à la phalloïdine. Magyar: Fluoreszcenciamikroszkópos felvétel marha tüdőartéria endotélsejtjeiről (Molecular Probes FluoCells prepared slide #2 (F14781)). A sejtmagok DAPI-val vannak festve (kék), a mikrotubulusokhoz anti-α-tubulin egéranitest, ahhoz pedig BODIPY FL-el jelölt anti-egér kecske-IgG van kapcsolva (zöld), míg az aktin filamentumok Texas Red-X-el kapcsolt falloidinnal vannak jelölve (vörös). A kép három felvétel szuperpozíciójával készült. Hamis színek. Lietuvių: Citoskeletas. Aktino filamentai – raudona, mikrovamzdeliai – žalia, branduolys – mėlyna spalva. Română: Sub microscop Celule endoteliale . microtubulii sunt de culoare verde, iar filamentele de actină sunt roşii, pe când nucleul celulei este colorat albastru Русский: Цитоскелет эукариот. Актиновые микрофиламенты окрашены в красный (фаллоидином, связанным с TRITC), микротрубочки — в зеленый (антителами, связанными с FITC), ядра клеток — в голубой цвет (DAPI). Клетки эндотелия лёгочной артерии быка. Українська: Цитоскелет еукаріот. Актинові мікрофіламенти забарвлені в червоний колір, мікротрубочки — в зелений, ядра кліток — в блакитний
context_page_description: An antibody, also known as an immunoglobulin, is a large, Y-shaped protein produced mainly by plasma cells that is used by the immune system to neutralize pathogens such as pathogenic bacteria and viruses. The antibody recognizes a unique molecule of the pathogen, called an antigen, via the fragment antigen-binding variable region. Each tip of the "Y" of an antibody contains a paratope that is specific for one particular epitope on an antigen, allowing these two structures to bind together with precision. Using this binding mechanism, an antibody can tag a microbe or an infected cell for attack by other parts of the immune system, or can neutralize its target directly. Depending on the antigen, the binding may impede the biological process causing the disease or may activate macrophages to destroy the foreign substance. The ability of an antibody to communicate with the other components of the immune system is mediated via its Fc region, which contains a conserved glycosylation site involved in these interactions. The production of antibodies is the main function of the humoral immune system.
context_section_description: Specific antibodies are produced by injecting an antigen into a mammal, such as a mouse, rat, rabbit, goat, sheep, or horse for large quantities of antibody. Blood isolated from these animals contains polyclonal antibodies—multiple antibodies that bind to the same antigen—in the serum, which can now be called antiserum. Antigens are also injected into chickens for generation of polyclonal antibodies in egg yolk. To obtain antibody that is specific for a single epitope of an antigen, antibody-secreting lymphocytes are isolated from the animal and immortalized by fusing them with a cancer cell line. The fused cells are called hybridomas, and will continually grow and secrete antibody in culture. Single hybridoma cells are isolated by dilution cloning to generate cell clones that all produce the same antibody; these antibodies are called monoclonal antibodies. Polyclonal and monoclonal antibodies are often purified using Protein A/G or antigen-affinity chromatography.
  In research, purified antibodies are used in many applications. Antibodies for research applications can be found directly from antibody suppliers, or through use of a specialist search engine. Research antibodies are most commonly used to identify and locate intracellular and extracellular proteins. Antibodies are used in flow cytometry to differentiate cell types by the proteins they express; different types of cell express different combinations of cluster of differentiation molecules on their surface, and produce different intracellular and secretable proteins. They are also used in immunoprecipitation to separate proteins and anything bound to them (co-immunoprecipitation) from other molecules in a cell lysate, in Western blot analyses to identify proteins separated by electrophoresis, and in immunohistochemistry or immunofluorescence to examine protein expression in tissue sections or to locate proteins within cells with the assistance of a microscope. Proteins can also be detected and quantified with antibodies, using ELISA and ELISpot techniques.
  Antibodies used in research are some of the most powerful, yet most problematic reagents with a tremendous number of factors that must be controlled in any experiment including cross reactivity, or the antibody recognizing multiple epitopes and affinity, which can vary widely depending on experimental conditions such as pH, solvent, state of tissue etc. Multiple attempts have been made to improve both the way that researchers validate antibodies and ways in which they report on antibodies. Researchers using antibodies in their work need to record them correctly in order to allow their research to be reproducible (and therefore tested, and qualified by other researchers). Less than half of research antibodies referenced in academic papers can be easily identified. Papers published in F1000 in 2014 and 2015 provide researchers with a guide for reporting research antibody use. The RRID paper, is co-published in 4 journals that implemented the RRIDs Standard for research resource citation, which draws data from the antibodyregistry.org as the source of antibody identifiers (see also group at Force11).
caption: Immunofluorescence image of the eukaryotic cytoskeleton. Microtubules as shown in green, are marked by an antibody conjugated to a green fluorescing molecule, FITC.
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