Flag tag protein detection antibody experimental application note

Flag tag protein detection antibody

Abbkine offers Flag antibodies for WB, IF, and IP applications, specifically for Flag tag fusion proteins, and Flag tag antibodies for recognition of Flag-tagged recombinant proteins expressed in cells, including Flag-based recombinant proteins at the amino terminus, mid-end, and carboxy terminus. .

The Flag tag system utilizes a short hydrophilic octapeptide (DYKDDDDK) to fuse to the target protein. The Flag tag can be located at the C-terminus or N-terminus of the protein. This system has been widely used in many cell types such as bacteria, yeast and mammalian cells, and the corresponding Flag tag antibody is also widely used. Since the purification conditions of the Flag tag system are non-denaturing, all active fusion proteins can be purified. The Flag tag can be removed by the addition of enterokinase, which specifically recognizes the 5 amino acid residues at the C-terminus of the peptide sequence. Flag antibodies can be used to detect and express the expression of Flag-tagged fusion proteins, intracellular localization, and to purify, qualitatively or quantitatively detect Flag fusion-expressed proteins.

Since the Flag tag protein detects the hydrophilic character of the antibody , the Flag tag is often located on the surface of the fusion protein and is therefore relatively easy to be accessed and recognized by the antibody. Different Flag tag antibodies have different recognition and binding properties to the Flag tag. Abbkine provides a monoclonal antibody with clone number 1B10, which is one of the most specific and potent Flag antibodies on the market.

product name

App types

Item number

specification

Anti-DDDDK Tag Mouse Monoclonal Antibody (1B10)

WB, IF, IP

A02010

50ul, 200ul

1ml, 10ml

Anti-DDDDK Tag Rabbit Polyclonal Antibody

WB

A02011

50ul, 200ul

1ml, 10ml

HRP Conjugated Anti-DDDDK Tag Mouse Mab (1B10)

WB

A02015

50ul, 200ul

1ml, 10ml

Fig. 1. Flag tag protein IP assay, IP (1:200) - WB (1:5,000): untransfected 293 cell lysate (lane A), 293 cell transfected lysate transfected with Flag tagged protein (lane B), using mouse IgG as a negative control immunoprecipitation 293 cell lysate (lane C), using Flag monoclonal antibody (1B10) IP transfected 293 cell lysate (lane D), 293 cell lysate Join only Protein G Beads (lane E).

Fig. 2. Using the Flag tag monoclonal antibody, the location of the transfected Flag recombinant protein in 293 cells was analyzed by immunofluorescence assay (1:2000). The secondary antibody was IFKine? Red é©´ anti-mouse, blue for DAPI Stained nuclei.

Abbkine is committed to providing the latest products and technologies for life science research and medical development worldwide. Abbkine designs, develops, manufactures and markets a range of unique products, including antibodies, research reagents and cell functional analysis products related to life science research. Their products provide effective solutions to the challenges of scientific research, saving time and increasing research efficiency for scientists around the world. As the general agent of Abbkine in China, Aijiejie Technology Co., Ltd., with its rich professional experience, will continue to provide Abbkine quality products and technical services to customers in China.

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