Lieke Widowati

6 Questions 7 Answers 0 Followers

Questions related from Lieke Widowati

His column crack?

Hi... I seem to be in a very big trouble. My His columns look cracked after my last purification. In another column, the materials seem to be pushed. How is it possible and is there anything I...

25 June 2024 7,030 2 View

High Pressure on ÄKTA Start without column?

Hi everyone, I really don't know how to fix the issue that I am facing. For background, I am working on the purification of inclusion bodies protein, which require the use of 8 M urea in the...

10 June 2024 9,465 0 View

Sample with 8M urea causes high pressure during IMAC?

Hi all, I have a problem with the high pressure of my HisPrep column. So these are the steps did before I loaded the sample: 1. Frozen cell pellet was resolubilized directly with lysis buffer...

13 May 2024 1,105 2 View

Any advice on how to do cell lysis using osmotic shock?

Hello everyone, I am supervising a master thesis student whose goal is to compare different lysis methods for recombinant protein purification. We are working with E. coli pellet and he is...

09 April 2024 6,211 7 View

Has anyone ever done protein refolding using affinity chromatography?

HI! Has anyone ever done protein refolding using affinity chromatography? My recombinant protein has a His Tag and usually to purify it we use IMAC on the ÄKTA machine. However the protein is...

03 March 2024 4,709 11 View

What is the best refolding methods you have worked with?

Hi everyone, I have never worked with Inclusion Bodies so do not have a method that really works. My protein is recombinant ß-casein that is produced both in the soluble fractions of the pellet...

23 February 2024 3,782 5 View