How to purify Proteins using Batch Spin

Protein purification is easier than imagined! Let us show you how to purify a protein using batch spin columns and affinity resin.

The full protocol including all mentioned buffers can be found here:
https://cube-biotech.com/products/pro...

The given example uses His-tagged GFP and our in-house developed Ni-INDIGO resin:
https://cube-biotech.com/products/pro...

The recipes for all mentioned buffers can be found here:
https://cube-biotech.com/media/ae/37/...

Other products of Cube Biotech that might interest you include:

His-Cube Ni-INDIGO His-tag protein purification Kits:
https://cube-biotech.com/hiscube-ni-i...

His-tagged eGFP (shown in the video):
https://cube-biotech.com/products/act...

Other His-tag purification products by Cube Biotech:
https://cube-biotech.com/products/pro...

Time stamps of the shown protocol:

0:00 - Intro
0:11 - Introduction
0:22 - Removal of storage buffer
0:43 - Preparation of protein binding
1:11 - Protein Binding
1:34 - Removal of unwanted cell lysate
1:47 - Washing
2:30 - Protein Elution
3:21 - Endcard

Individual steps of the protocol:
https://cube-biotech.com/Batch-Spin-H...

1. Pipette 1 ml Ni-INDIGO agarose slurry into a reaction tube.

2. Centrifuge the agarose particles at 500 x g. Then remove the storage buffer on top.

3. Add binding buffer to the agarose. The recipes for all required buffers can be found in the description below.

4. Shake the reaction tube.

5. Remove excess binding buffer by centrifuging at 500 xG. We recommend to repeat the two previous steps (3 and 4 once).

6. Now add the His-tagged protein lysate to the reaction tube.

7. Let the mixture incubate in the fridge shaking for one hour at 4°C.

8. Centrifuge the mixture at 500 x g.

9. Remove the excess protein lysate.

10. Add wash buffer.

11. Let the mixture incubate in the fridge shaking for one hour at 4°C.

12. Centrifuge the mixture at 500 x g. Then remove the wash buffer on top.

13. Repeat the washing procedure (Steps 10-12) for 4 times.

14. Add 1 ml Elution Buffer.

15. Let the mixture incubate in the fridge shaking at least 10 minutes at 4°C.

16. Centrifuge the elution buffer and the agarose at 500 x g. We recommend to repeat this and the two previous steps at least twice.

17. Pipette the elution buffer on top into a new reaction tube.

18. The new reaction tube now contains your purified His-tagged protein.