Simi larly, epi tope tag ging can allow for tracking closel y relate d protein s without fear of spurious resul ts resulting from cross -reac tive antibodies. Experiment s using antibodies against epitopes foun d in the native molecule cannot provide a comparable negative cont rol. Background : Belongs to the histone H2B family. Immunohistochemistry analysis of paraffin-embedded Human Breastcancer using Histone H2B antibody.High-pressure and temperature Sodium Citrate pH 6.0 was used for antigen retrieval.
For example, because the tag would be missi ng from extract s of cel ls that are not expressing a tagg ed protein, negative controls are unequivocal. Western blot analysis of Histone H2B in HeLa lysates using Histone H2B antibody. However, epitope tagging offers a numbe r of additional advanta ges. The most obvious advantage of epitope tagging is that the time and expense associated with gene rating and characte rizing antibodies against multiple proteins are obviated. High ly spec ifi c antibodies and useful cloning vector s encoding epitope tags adjacent to cloning site s are readi ly available from commercia l suppli ers or erstwhi le collaborators, adding to the ease of initiat ing such studies. This stands in sharp contrast to the several months that would otherwise be spent generat ing and character izing antisera against the pro- tein itsel f. Part 4 overview 4.1 Western blot troubleshooting 4.2 IHC and ICC troubleshooting 4.3 Flow cytometry troubleshooting 4.1 Western blot troubleshooting This detailed guide takes you through some of the most common problems, like no signal, high background and multiple bands, and what you can do to fix them. Tagging a protein with an existi ng epitope is a simple procedure that allow s researcher s to readi ly pu rify or follow proteins throug h meaningfu l and revealing experiments quite promptly after expressing a cloned se- quenc e.
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