BG GST FF Affinity Chromatography Resins

BG GST FF Affinity Chromatography Resins are designed for the purification of GST-tagged fusion proteins. The GST tag is widely used in research projects for its ability to enhance the solubility of recombinant proteins. This product uses glutathione as a ligand to achieve specific affinity binding with GST, allowing for rapid extraction of GST-tagged fusion proteins from cell fermentation broth.

2.Features

Chromatographic operation Procedure

BG GST FF Affinity Chromatography Resins are affinity chromatography products widely used for the separation and purification of glutathione S-transferase. The process typically includes equilibrium, sample loading, washing, and elution steps. The specific operating procedures are as follows:

1.      Equilibration: Equilibrate the column with 5-10 column volumes (CV) of equilibration buffer until the conductivity and pH of the effluent remain constant (consistent with the equilibration buffer). The typical equilibration buffer is 50 mM Tris-HCl, pH 8.0.

2.      Sample Loading: The sample buffer should be as consistent as possible with the equilibration buffer. Solid samples can be dissolved and prepared in equilibration buffer; dilute sample solutions can be dialyzed with equilibration buffer; concentrated sample solutions can be diluted with equilibration buffer. To avoid clogging the chromatography column, samples should be centrifuged or microfiltered. The sample loading volume should be calculated based on the resins' binding capacity and the target protein content in the sample.

3.      Washing: After loading, continue washing with equilibration buffer until the UV absorbance drops to baseline.

4.      Elution: The elution buffer is typically 50 mM Tris-HCl + 10 mM GSH, pH 8.0 (GSH is easily oxidized, prepare fresh as needed).

5.      Regeneration: After multiple uses (depending on the fermentation broth sample and impurity characteristics), regeneration is required.

(1) Clean with buffer 1: 0.1 M Tris HCl + 0.5 M NaCl, pH 8.5 for 2 CV;

(2) Clean with buffer 2: 0.1 M sodium acetate + 0.5 M NaCl, pH 4.5 for 2 CV;

(3) Repeat the above steps 3-4 times for a total of 4-5 washing cycles with alternating buffers;

(4) Re-equilibrate with 3-5 CV of 1×PBS.

If the gel seems to lose binding capacity, it may be due to precipitation, denatured proteins, or accumulation of non-specifically bound proteins. To remove precipitated or denatured substances, clean the resins with 2 column volumes of 6M guanidine hydrochloride, followed immediately by 5 CV of 1×PBS. To remove hydrophobically bound substances, clean with 3-4 CV of 70% ethanol or 2 column volumes of non-ionic detergent (0.1% concentration), followed immediately by 5 CV of 1×PBS.

Note: For long-term storage (> 1 month), use the following additional cleaning procedure:

(1) Clean the gel twice with 10 bed volumes of 1×PBS;

(2) Repeat cleaning with 20% ethanol.

6.      Storage: Store at 2-10°C in 20% ethanol solution.

7.      Other considerations: Avoid column dry-out or loose sealing during use and storage to prevent air bubbles from entering.

3.Specification