Why does the column turn brown?

The column will only turn brown if there is a reducing agent such as DTT or beta-mercaptoethanol (BME) in the buffer(s). The reducing agent complexes with the cobalt ion to form a brown precipitate. Using up to 10 mM BME should not affect the performance of the resin significantly. If BME is not used, the resin has turned brown, and there is no protein binding, DTT or DTE may be present in the [...]

What should I do if purified protein fails to elute from TALON resin?

Check to see if any of the following factors are responsible and try the recommended solutions listed below: 1. Is the protein being expressed? Check the lysate using Western blotting, preferably with a 6xhis antibody. This also assures that the reading frame is the same for the protein and the tag. 2. How does the lysate preparation look? Was the lysate preparation turbid? If not, the cells [...]

What proteins have been extracted with xTractor Buffer at Takara Bio, and from what types of cells?

We have tested xTractor Buffer on various E. coli host strains expressing the following proteins: 6x(HN)-LacZ (beta-galactosidase), 6x(His)-LacZ, 6x(HN)-GFPuv (Green Fluorescent Protein), 6x(His)-GFPuv, HAT-DHFR (DiHydro Folate Reductase), and 6x(HN)-Luciferase. Both beta-galactosidase and luciferase were functionally tested following purification and were found to retain their enzymatic [...]

What is the effect on protein yield if xTractor Buffer is used with a fresh bacterial pellet compared to a frozen cell pellet?

We have not seen a significant difference in yield when using the xTractor Buffer with freshly collected bacterial pellets and frozen pellets.Protein purification products Care guide Caution [body] This care sheet provides general information only for handling Carolina™ bacterial cultures. When you work with bacteria, it is imperative that you use sterile techniques at all times. Failing to [...]

What are the components of xTractor Buffer?

The components of xTractor Buffer are proprietary. However, the buffer does not contain chelating agents or protease inhibitors and is fully compatible with TALON resins and spin columns for purification of his-tagged proteins.Protein purification [...]