Epitope tagging has become a powerful tool for detection and purification of expressed proteins. Epitope tags are short peptide sequences that are easily recognized by tag-specific antibodies. Due to their small size, epitope tags do not affect the tagged protein’s biochemical properties. Most often sequences encoding the epitope tag are included with target DNA at the time of cloning to produce fusion proteins containing the epitope tag sequence.
Anti-epitope tag antibodies are useful for identification, immunoprecipitation or immunoaffinity-purification of a recombinant protein. Anti-FLAG (YKDDDDK)-tag polyclonal antibody was raised by immunizing a rabbit with the peptide YKDDDDK conjugated to KLH.
- Western blot dilution: 1/2,000
- ELISA (assay dependent)
Other applications have not been tested.
Immunogen: DYKDDDDK cross-linked to KLH
Specificity: This antibody recognizes FLAG-tagged fusion proteins. However, this antibody does not react with “ so called yeast Flag Tag “, consisting of three or nine repeats of DYKDHD sandwiched with G.
Form: Antiserum with 0.05% sodium azide
Storage: Shipped at 4°C and long term storage -20°C
- Brizzard BL et al “Immunoaffinity purification of FLAG epitope-tagged
bacterial alkaline phosphatase using a novel monoclonal antibody and peptide
elution.” Bio Techniques 16: 730-735 (1994) PMID: 8024796
Fig. 1 Detection of FLAG-tagged protein with this antibody by Western blot.
-: Lysate of 293T cells transfected with an empty vector
+: Lysate of 293T cells transfected with the plasmid carrying the FLAG-tagged PRMT6 gene