Epitope tags and anti-epitope tag antibodies are widely used for purification and detection of recombinant proteins in immunoblotting, immunostaining and immunoprecipitation.Epitope tagging is widely used in the characterization of newly discovered proteins. With the development of biological technology, researchers can by recombinant DNA techniques, to construct containing the target gene and epitope tagged fusion protein, and then through the specific tag antibody on the identification and purification, in order to achieve the research needs.The major categories:Many different epitope tags have been engineered into recombinant proteins. Sino Biological Inc. has antibodies against FLAG®( DYKDDDDK), HA, HIS-tag, c-Myc, GFP, GST, V5, S-tag, T7-tag, E-tag and Glu-Glu tag.
Tag antibody is a special area or group that determines the specificity of the antigen molecule, and is the structure or sequence of the antibody specific binding. With the development of biological technology, researchers can construct the fusion protein containing target gene and epitope tagged by DNA recombination technology, and then identify and purify the specific tag antibody to meet the requirements of the research.
S-tag is often used as tag in N terminal or C terminal in protein and expressed with protein. Although S-tag is not the most commonly use as antibodies against His or GST, but it is still widely used and recognized in the field of biological research and related fields. S-tag has sequence of KETAAAKFERQHMDS. Antibody against S-tag is often been used in the detection of WB and immunofluorescence and often been used to study the expression of proteins and the interaction between proteins and proteins.
Immunofluorescence staining of S-Tag in Hela cells, transfected with pSTEP2-Stag-GFERh (Figure A), pSTEP2-his-GFER-Stag (Figure B), pSTEP2-his-Stag-GFER (Figure C), pSTEP2-Stag-GSTT2h (Figure D),pSTEP2-his-Stag-GSTT2 (Figure E) and pSTEP2-his-GSTT2-Stag (Figure F). Cells were fixed with 4% PFA, permeabilzed with 0.3% Triton X-100 in PBS, blocked with 10% serum, and incubated with Mouse anti-S-Tag monoclonal antibody at 37℃ 1 hour. Then cells were stained with the Alexa Fluor® 488-conjugated Goat Anti-mouse IgG secondary antibody (green) and counterstained with DAPI (blue).