Philipp Czermak

7 Questions 26 Answers 0 Followers

Questions related from Philipp Czermak

Why is my protein eluting in the salt wash when applying IMAC?

Hi everyone! I currently have the issue that my proteins are all eluting at the salt wash that I apply during my IMAC (HisTrap 5ml HP column). My proteins are all His-SUMO-tagged. They are...

26 May 2024 8,396 3 View

Serum-free medium or medium sublemented with serum for Sf9 protein expression? What are the advantages? Is any BEVS superior to the other?

Hi everybody! I am currently facing a very nasty protein that I am trying to purify. Besides the need for several purification steps, the lack of proper expression in insect cells gives me a hard...

03 February 2022 4,203 0 View

Why is my protein not binding to my reversed-phase column?

Hello everyone! Short facts: I am trying to perform reversed-phase chromatography via a desalting column (Waters: Sep-Pak tC18 3cc) of an already purified hydrophobic protein to exchange the...

02 February 2021 5,818 15 View

Does anyone have experience with small size exclusion columns on an FPLC (NGC Bio-Rad)?

Hi, We are running and validating some interactions of proteins via running them over a size exclusion column. We want to switch to a smaller one in order to save protein. Currently, we have a...

11 May 2020 3,617 2 View

In vitro binding assay: Why do my karyopherins do not bind cargo?

Hello everyone, I am trying to decipher the interaction between different karyopherins (importin alpha and importin beta) and we also want to show them binding to an NLS containing cargo, to...

30 April 2020 1,184 7 View

Lyophilization troubleshooting of nucleoporin

Hi all, I purified, performed Reversed-Phase Chromatography (Strata-SDBL columns), and lyophilized my nucleoporin (truncated version with mainly FG-Domains) several times already. Until now I...

01 January 1970 5,128 6 View

No Phenotype upon knock-out

Hello everyone, Right away: This is a question I am struggling with, and it is asked for a PhD program that I want to get into. So maybe I am too stupid and shouldn’t participate since I can’t...

01 January 1970 4,827 5 View