Speed up your His-tag Protein Detection

The proflo His-tag quick test is a competitive immunochromatographic lateral flow assay which allows quick and easy detection of your histidine-tagged proteins. Thus, it represents a fast method to monitor His-tag proteins in small-scale experiments, during your scale-up procedure as well as during your actual purification process. The semi-quantitative assay is suitable for determination of His-tag proteins from cell lysates as well as purified recombinant protein preparations with a concentration range of 2 - 50 µg/ml (for a 25 kDa protein). 

Applications

  • expression monitoring of recombinant His-tag proteins
  • for small-scale screening
  • during expression
  • during scale-up
  • during the purification process

 

Benefits

  • quick results in only 10 min
  • easy handling - 2-step protocol
  • faster than SDS-PAGE and Western Blot
  • works in small sample volumes
  • robust and reliable, similar to WB & ELISA
  • reproducible data

proflo His-tag Quick Test - Test Principle

The proflo His-tag quick test is a competitive immunochromatographic lateral flow assay containing a specific anti-His-tag antibody bound to gold particles (capture antibody complex), immobilized His-tag protein (test line) and a control antibody (control line). After application of the sample, the capture antibody complex flows by capillary forces across the strip membrane. With increasing amount of recombinant His-tagged protein in the sample, the intensity of the test line will decrease whereas the control line is independent of the amount of recombinant His-tagged protein and should always appear with the same intensity.

Test line: high intensity

  • the capture antibody complexes remain unbound
  • at the test line: capture antibody complexes bind to the immobilized His-tag protein
  • at the control line: excessive capture antibody complexes are captured by the control antibody

Result: no His-tag protein in the tested sample.

Test line: low intensity

  • the capture antibody complexes bind the recombinant His-tag protein in the sample
  • at the test line: unbound capture antibody complexes bind the immobilized His-tag protein
  • at the control line: capture antibody complexes bound to the recombinant His-tag protein are captured by the control antibody

Result: intermediate amount of His-tag protein in the tested sample.

Test line: hardly visible

  • the capture antibody complexes bind the recombinant His-tag protein in the sample
  • at the test line: less capture antibody complexes are bound to the immobilized His-tag protein
  • at the control line: capture antibody complexes bound to the recombinant His-tag protein are captured by the control antibody

Result: high amount of His-tag protein in the tested sample.