Lieke Widowati

10 Questions 8 Answers 0 Followers

Questions related from Lieke Widowati

Any way to solubilize inclusion bodies without Urea or Gua HCl?

Hi everyone, I have a very troubling question with the project I'm working on. So just a brief overview: we want to produce a eukaryotic recombinantly, the host is E. coli. Production is done is...

22 October 2024 6,455 1 View

Line on the bottom of SDS-PAGE gel?

Hi smart people! I have a small curious question. Attached are my student's gel - which looks weird as there is always a line on the bottom (near the smallest marker). This is self-made gels...

27 June 2024 9,581 1 View

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 7,109 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 9,611 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 1,287 2 View

SDS PAGE gel staining not work?

Hi everyone, I have a question that has been troubling the whole lab here with the SDS-PAGE gel staining. We usually use colloidal gel staining (ROTI blue quick) and it works fine every time....

23 April 2024 5,737 0 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 6,290 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 4,792 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 3,843 5 View

What is the solubility of casein in urea solution?

Hi everyone, Does anyone happen to know the solubility of casein in urea solution? I have a problem with the purification of recombinant casein as most of the protein are produced in inclusion...

22 February 2024 4,937 3 View