I have a vector pET23b with monomeric EGFP-6X His tagged cloned in that. The plasmid is transformed in BL21(DE3) cells. Kindly suggest me some good literature for the purification of the GFP-His tagged protein.
Hello, here same topic discussed here
https://www.google.com/amp/s/www.researchgate.net/post/Can_anyone_share_his_her_experience_purifying_GFP_or_GFP_variants_in_e-coli/amp
Hope this info will help you.
Good luck!
Hello Puneeta Bathla!
I have read some topics on the internet that you can use His-Tag affinity columns or also GFP affinity columns. They said that the second method is cleaner and leads to a higher yield than using the first one, although it seems to be expensive. Here are some links and protocols that might be useful on these topic: https://www.chromotek.com/products/nano-traps/gfp-trap/
Article Purification of GFP fusion proteins with high purity and yie...
Also using the GFP interaction, a new article describe a method of purification using anti-GFP/YFP nanobody sepharose columns:
Article Selectable High-Yield Recombinant Protein Production in Huma...
If you want to try first with common His-Tag affinity columns you can use Ni-NTA columns and there are also some articles on the literature and also here on RG on this topic:
http://www.cell.com/structure/pdf/S0969-2126(14)00115-4.pdf
https://www.researchgate.net/post/Can_anyone_share_his_her_experience_purifying_GFP_or_GFP_variants_in_e-coli
I am not a specialist on this topic but I hope this can help you!
Good Luck!
Hello, here same topic discussed here
https://www.google.com/amp/s/www.researchgate.net/post/Can_anyone_share_his_her_experience_purifying_GFP_or_GFP_variants_in_e-coli/amp
Hope this info will help you.
Good luck!
Check these articles
Article Cloning, expression and two-step purification of recombinant...
https://www.med.upenn.edu/robertsonlab/assets/user-content/documents/Roche%20NiNTA%20protocol.pdf.pdf
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