Question

Why do we use an epitope tag versus a direct antibody for our protein of interest? How you would go about making an epitope tag for your favorite gene of interest?

Answer 1

Epitope Tags and anti-tag antibodies are valuable for various immunoassays such as western blotting,flow cytometry etc.Epitope tags are the most efficient approach to detect newly found proteins for which specific antibodies are not yet available or for targets that are less immunogenic.

moreover,epitopetags are helpful for

1. affinity purification of target proteins
2. improving solvency of target proteins
3.securing against intracellular cleavage of protease.
4.providing enzymatic function for target protein

*making epitope tag for favourite gene of interest:

In genomic epitope labeling for favourite gene, at first make an epitope tag as smaller than normal exon flanked by splice donor and acceptor locales for inserting into the intron of a plasmid clone of a target gene. The tag is named the CD tag after the "central dogma" of science on the grounds that the target gene is tagged at first, trailed by the communicated mRNA, lastly, after completion of translation, the expressed protein. The method was later improved by the utilization of transposons for introducing into the target gene