GST and 6HIS Tag
The production of fusion proteins requires the confirmation of their structural integrity and the accessibility of the associated tag following isolation. The GST and 6HIS check kits are simple tools for assessing tagged proteins during production processes and drug discovery.
- Ready-to-use kits
- Simple and rapid assay protocols
- Amenable to miniaturization
- Compatible with a variety of buffers and media
- Measurement of GST or 6HIS proteins
- Monitoring of GST and 6HIS tag accessibility in fusion proteins
- Elimination of HTS false positives due to compound-tag interference
Assay Principle:
The GST and 6HIS check kits are competitive immunoassays that use Cryptate-labeled anti-tag antibodies and XL665-labeled tags. The check kit assays can be run in a single microplate requiring only a two hour incubation. Because of the ratiometric measurement of HTRF®, tagged proteins can be accurately detected in a variety of buffers and media.
Product Specifications:
Protein Quantification:
A fast and simple HTRF protocol makes it possible to measure GST or 6-HIS tagged proteins at each step of
protein production and purification. These kits can be calibrated using the appropriate protein, making
the assay very accurate and compatible with the SOPs used in industrial processes.
Drug Discovery:
6HIS and GST check kits are useful after HTS to eliminate false positives. Compounds that interrupt the
binding of tag-detection reagents (e.g. HTRF® MAb Anti-6HIS-Cryptate) will show strong signal inhibition
in the check kit. These compounds can be removed from further charactarization, saving time and
resources.
Tag Accessibility:
The 6HIS check kit is a useful tool to determine whether the tag is accessible to antibodies and
to predict the possibility of purifying the protein using a Nickel column. If a 6HIS fusion protein is
not detected using our check kit then the tag is inaccessible and not available for purification or other
binding purposes.
| GST tag check kit | |
| Detection limit: | 1 ng/mL (average) |
| S/B= | 15 in 96- or 384-well formats |
Figure 1: A standard curve using native GST was performed following the protocols detailed in the product inserts. The detection limit of this assay was 0.26 ng/ml
Assay Performance:
The GST check kit enables the verification of tag accessibility on GST-tagged fusion proteins. Figure 2 shows the dose response curves of different GST fusion proteins assessed using this kit. All six GST containing proteins were detected using our competitive immunoassay.
Figure 2: GST check kit was performed using the standard protocol and a series of six different GST proteins.
Ordering Info:
Check Kit Assays:
| Description | Quantity* | Cat no. | Product Insert |
| 6HIS tag check kit | 1,000 tests | 62HISPEB | |
| GST tag check kit | 1,000 tests | 62GSTPEB | |
| *based on 20 µl assay volume | |||
For more information or To Place an Order:
U.S. and Canada
1-888-963-4567
Europe and other countries
+33(0)466 796 705
Japan
+81-3-5510-2932
