|Tested applications||WB IHC IF IP|
|Recommended Dilution||WB 1:2000 - 1:5000
IHC 1:50 - 1:500
IF 1:50 - 1:500
IP 1:50 - 1:100|
|Observed MW||Refer to Figures|
|Immunogen||A synthetic Peptide of human DDDDK-Tag|
|Storage Buffer||Store at -20℃. Avoid freeze / thaw cycles.
Buffer: PBS with 0.02% sodium azide, 50% glycerol, pH7.3.|
Western blot analysis of over-expressed DDDDK-tagged protein in 293T cell using DDDDK antibody at different dilution. Each lane was loaded with 2 ug cell lysate.
Immunoprocipitation of over-expressed DDDDK-tagged protein in HeLa cell incubated with DDDDK antibody. A mock served as negative control and over-expressed Hela cell lysate served as positive control.
Protein tags are protein or peptide sequences located either on the C- or N- terminal of the target protein, which facilitates one or several of the following characteristics: solubility, detection, purification, localization and expression. The DYKDDDDK(FLAG) peptide has been used extensively as a general tag in expression vectors. This peptide can be expressed and detected with the protein of interest as an amino-terminal or carboxy-terminal fusion. N-terminal FLAG vectors provide an Ek cleavage site for removal of the fusion tag. The FLAG peptide is likely to be located on the surface of a fusion protein because of its hydrophilic nature. As a result, the FLAG peptide is more likely to be accessible to antibodies. A DDDDK-tag can be used in many different assays that require recognition by an antibody, such as western blotting, immunocytochemistry, immunoprecipitation, flow cytometry, protein purification, and in the study of protein-protein interactions, cell ultrastructure, and protein localization and so on. This antibody is a mouse monoclonal antibody raised against 3xFlag (3xDYKDDDDKT) sequence and recognizes the (3x)DYKDDDDK peptide and detects DDDDK-tagged proteins.
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