His-Tag Quick Test, BioAssay(TM)

Product Overview

• Article Name: His-Tag Quick Test, BioAssay(TM)
• Biozol Catalog Number: USB-550799
• Supplier Catalog Number: 550799
• Alternative Catalog Number: USB-550799-25
• Manufacturer: US Biological
• Category: Kits/Assays

Product Description

For purifying recombinant proteins, the histidine tag (His-Tag) is the most widely used peptide tag with usually 6-10 consecutive histidines at the N- or C-terminus of a target protein. Purification of His-tagged proteins is done by using immobilized metal ions like nickel or cobalt in a variety of different buffer conditions. The His-Tag Quick Test, BioAssay(TM) 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 2-50ug/ml (for a 25kD protein). 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 Western Blot and ELISA Reproducible data Test Principle: The test is based on a competitive reaction between a specific anti-His-tag antibody bound to gold particles (capture antibody complex) localized at the bottom of the test strip and a His-tagged protein immobilized on the test line of the test strip. After applying a sample to the test strip, the capture antibody complex binds the His-tagged recombinant protein present in the sample and flows by capillary force across the strip membrane. Capture antibody complexes that are not bound to recombinant proteins are captured by the His-tagged proteins at the test line. Capture antibody complexes that are not captured by the His-tagged proteins at the test line bind to specific control antibodies located at the control line. Thus, with increasing amount of His-tagged protein in the sample, the test line staining will appear weaker. If the test line is not visible due to saturation of binding sites on the capture antibody complexes, a new test strip should be used with a higher dilution of the sample if analysis or comparison of samples with higher concentrations is intended. Depending on the size of your protein and accessibility of the His-tag, the intensity of the test line can vary significantly. Note: The control line is not dependent on the amount of His-tagged protein in your sample and should always appear as a strong purple line. Without a clearly visible control line, the test is not valid. The binding to the control antibody could be inhibited in such cases. The test should be repeated with a higher dilution of the sample. Kit Components: Test strips, 25x His-tag lateral flow test strips Buffer Dilution buffer, 2x2ml Reaction tubes, 25x2ml Storage and Stability: Store the test kit and components at 4C. Unopened kit is stable for 6 months after receipt.