The Flag Check kit enables the rapid detection of Flag tagged fusion proteins.
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The Flag Check kit enables the rapid detection of Flag tagged fusion proteins.
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Overview

The kit enables the rapid detection of Flag tagged fusion proteins at each step of protein production and purification. It can either be used to ensure that the Flag motif of a fusion protein is accessible to the anti-Flag antibody, or to approximate the concentration of a Flag-tagged protein. Calibrated using the appropriate protein, the assay is very accurate and compatible with the SOPs. It can also be used to eliminate false positives.

Benefits

  • ELIMINATION OF FALSE POSITIVES
  • PROTEIN QUANTIFICATION
  • TAG ACCESSIBILITY

Assay principle

As shown in the graph, Flag peptide labeled with d2 is detected by anti Flag Cryptate conjugate (anti-tag Eu-Cryptate). The Flag-tagged fusion protein (Tag fusion protein) to be ascertained competes with the Flag-peptide-d2 conjugate (Tag-d2) for binding to the anti-Flag-Cryptate conjugate. The specific signal is inversely proportional to the concentration of Flag-tagged fusion protein.
assay principle of HTRF Flag Check kit

Assay protocol

The example on the right describes the protocol using a 20 µL final assay volume. Dispense 10 µL of standard or of Flag-tagged protein of interest into a well, with 5 µL of Flag Cryptate conjugate and 5 µL of Flag-d2 conjugate. Incubate for 2 hours at RT and read.
FLAG check kit assay protocol

FLAG Check Kit performance

The Flag Check Kit enables the rapid detection of FLAG-tagged fusion proteins. It can either be used to ensure that the Flag motif of a fusion protein is accessible to the anti-FLAG antibody, or to approximate the concentration of a FLAG-tagged protein.
Detection of Flag tagged-protein with the HTRF Flag check kit
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