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.
DYKDDDDK tag is known as Flag® tag which is often used as tag in N terminal or C terminal in protein and expressed with protein. (Anti-FLAG® is a registered trademark of Sigma-Aldrich Biotechnology.) DYKDDDDK tag is one of most commonly used in research of protein expression and protein interaction. Antibody against of DYKDDDDK tag is one most important epitope antibody for the detection of tag in Western Blot, immunofluorescence IF / ICC and Flow Cytometry / FACS. Additionally, DYKDDDDK tag is often used to purification of protein with DYKDDDDK tag (Flag® tag) in N terminal or C terminal of fusion protein. Antibody against DYKDDDDK tag (Flag® tag) has been widely used for purification of proteins which been expressed in bacteria, yeast and mammalian cells.
Immunofluorescence staining of DYKDDDDK tag in 293 cells, transfected with PCMV-CDH1-flag (Figure A), pSTEP2-Flag-FABP4-GST (Figure B), pSTEP2-Flag-ARG1-GST (Figure C), Flag-PRMT5-His (Figure D),Flag-PRMT6-His (Figure E) and His-flag-CD38(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-flag-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).