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airie 11-09-2009 02:26 AM

Applications for antibodies
Hello everyone, I am looking for primary antibodies for immunohistochemistry on PFA-perfused, cryoprotected frozen sections.
A fair few of the antibodies list different applications like Western Blotting, ELISAs, Immunoprecipitation and not Immunohistochemistry.

I feel really confused and out of my league, and do not understand why some of these antibodies cannot be used for an immunohistochemistry and immunocytochemistry? Are they conjugated to something else?!

Thank you for helping me clarify this....

Zagami 02-14-2010 11:30 AM

Re: Applications for antibodies
Otherwise from ELISA and WB methods, immunohistochemistry (IHC) it has the ability to show where certain proteins are located, inside a tissue section (normal or in relationship to its pathology), that deriving from a specific patient (age, sex, etc.), and effected biopsy at unfixed time. In IHC procedure, tissue section must be fixed before to prevent chemical or biological changes site of located proteins. Fixation process involves that proteins and other cellular components of the tissue, cannot stir anymore, to modify conformationally of it, to interact the one of it with the others, and this condition been termed “native molecules”. The produced antibodies against native molecules, in one conformational natural state (folded state), recognize only epitopes that has been present in this molecular form, while if the same proteins has been submitted to denaturation to get a linear molecules, such as WB during the preparation gel electrophoresis, the elicites antibodies cannot be able to recognize the same epitopes expressed on the native protein, still more if these are found in the fold (epitopic inaccessibility). Nevertheless, possibility that slightly denaturing tissue can to determine changes of native molecules with epitopes expression which are able to react with the antibody that detect same antigen in ELISA and WB methods. In add, two types of monospecific first antibodies to used in immunoassay : a) clonal, and b) monoclonal. The first recognize solely very specifically selected linear epitope on the antigen molecule after its detailed proteomic analysis, whereas the monoclonal antibodies recognize very often steric epitopes that frequently change their conformation during tissue preparation (denaturation tissue, protein extraction, protein transfer in the case of western blot – related procedures, or as a consequence of the protein-protein interactions, etc.) making the corresponding monoclonal antibody less specific and less avid. The various methods, ELISA, WB and IHC, seeking the same molecule, they use antibodies that not individualize the same epitope on molecule target, for different state (native or modified) of the inducer molecular antigen at correspondent antibody formation.

StephenJones01 09-26-2012 05:50 AM

Re: Applications for antibodies

Antibodies are substances found in the blood that help the body fight infection. They recognize foreign invaders and bind to chemicals on their surfaces called antigens. The process tags bacteria, viruses, infected cells or other foreign matter for destruction by other parts of the immune system. White blood cells called B-lymphocytes produce specific antibodies for every foreign antigen the body encounters. Antibodies are also referred to as immunoglobulins and are found in the blood. They are produced by white blood cells called B cells. Antibodies are one main defense of the immune system; they identify and sometimes deactivate bacteria and viral cells.

ELISA, or enzyme-linked immunosorbent assay, is used as a diagnostic tool in medicine and in biological research to quantify the amount of a specific protein. ELISA uses antibodies to detect the protein of interest and a reagent that produces a color. The amount of color relates to the amount of protein. Antibodies are part of the body defense against foreign proteins present on a bacteria or virus for instance. An antibody is specific for a single protein; this protein target is known as an antigen. For Immunohistochemistry, it is a powerful tool for diagnosing cancer and other diseases.

Best Regards,
Stephen Jones

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