FLAG Tag

Epitope tags have application in the labeling, isolation and detection of proteins using immunoblotting, Immunoprecipitation and immunostaining techniques. Epitope tags can be used, by affinity chromatography, to separate recombinant, overexpressed protein from wild-type protein expressed by the host organism. Due its small size the protein’s biochemical properties seem not to be affected by the tag protein. FLAG-tag, or FLAG octapeptide, a polypeptide consisting of eight amino acids (DYKDDDDK) can be added to a protein using recombinant DNA technology; therefore, the tag can be detected with the fused protein in many different assays that require recognition by an antibody.

The antibody was raised against DYKDDDDK (FLAG) synthetic peptide conjugated to KLH.

The antibody recognizes the N-terminal, C-terminal or internal DYKDDDDK-tagged fusion proteins.

FG4R

IgG2b

None

Purified with protein G, stored in PBS pH 7.4 and lyophilized.

If reconstituted with deionized water in 100 µl: WB 1:1000-3,000, IHC 1:500-2,000. Optimal dilution has to be determined by the user.

Research Use Only

Lyophilized antibodies can be kept at 4ºC for up to 3 months and should be kept at -20ºC for long-term storage (2 years). To avoid freeze-thaw cycles, reconstituted antibodies should be aliquoted before freezing for long-term (1 year) storage (-80ºC) or kept at 4ºC for short-term usage (2 months). For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made with the assay buffer. After the maximum long-term storage period (2 years lyophilized or 1 year reconstituted) antibodies should be tested in your assay with a standard sample to verify if you have noticed any decrease in their efficacy. To limit antibody loss or degradation, BSA (final concentration 1%) and sodium azide (final concentration 0.02%) can be added to the suggested first dilution. It is important to first verify if those preservatives are compatible with your assay.