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Blot and Gel Detection

Purification, characterization, and identification of proteins or nucleic acids often require analysis of the target in blotting applications. Proteins or nucleic acids are usually separated by size using gel electrophoresis and transferred to a membrane with a blotting technique.

A specific protein can be identified from a mixture using different detection methods. Primary antibodies can be used to recognize unique epitopes or genetically engineered fusion protein tags. Lectins with specificities to certain carbohydrate residues can be used to probe for a particular glycoprotein. These reagents can then be detected and visualized with an enzymatic or fluorescent detection method.

Nucleic acid blotting applications also require high specificity which is derived from the uniqueness of the target sequence and the fidelity of the complementary probe. The target sequence is detected by a complementary probe labeled with tags such as biotin, digoxigenin (DIG), DNP, or fluorescein. The label is then detected with an antibody to the tag and subsequent detection reagents.

Detection sensitivity for both protein and nucleic acid blotting applications depends on the abundance of the target and the quality of the reagents available for detecting the primary antibody or hybridized probe.