I am expressing the protein in the E. coli host cell, followed by the purification using dialysis buffer (20 mM phosphate buffer, 100 mM NaCl, 2 mM EDTA, 0.01% sodium azide, pH 7) for 12 h at room temperature by replacing old buffer with fresh after every 3 h.
As the literature suggests that EDTA is not a good choice for storing the protein in a dialysis buffer (as mentioned in the first paragraph), what would be the best way to store the purified protein?
Reference...
Article Drawbacks of Dialysis Procedures for Removal of EDTA
Dear Bhupendra Singh
if it true the EDTA removal could be difficult by dialisys, you can remove it quite well using desalting which is a sort of rapid SIZE exclusion suitable for buffer exchange.
in the following videos you can find more information about desalting:
https://www.blogger.com/video.g?token=AD6v5dzeX21U_Er6jBChQIhE5zWtQhNjRjNXRPR7qk7KfWWSQxhf8PbUn4R6KLL0pn_7tU92M0-B3kEUHO9IkTpelmdO79QR53uSg29ZFV3j5D-siHAamTu1y2wOEQ8S4As_wpxzLvA
https://www.blogger.com/video.g?token=AD6v5dwNlVHz3BclDT9RLY-6KbEEqkS3RnRdHn4BqTsZW4j_prf3KZwHDMTzRYT-B9Yifvmde7RdExL5AArLFtg1Q13fu4-83o3M8bCka1jJ5PfWFK8B53BOpx6YrzvLr2qDiPgT15c
however the best storage buffer depends from the protein properties. Why did you add EDTA? it was required for purification, to protect your protein from protease digestion? it is your protein a metal binding protein or it tend to aggregate in presence of metals?
best regards
Manuele
Yes. My protein aggregates in the presence of metal.
In general , Why researchers want to remove EDTA from their protein?
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