Mikel Irastortza-Olaziregi

6 Questions 13 Answers 0 Followers

Questions related from Mikel Irastortza-Olaziregi

In-resin MNase digestion, any tips?

I routinely perform pulldowns in order to purify a protein of interest, and afterwards I digest linear nucleic acids attached to my bait protein using micrococcal nuclease (MNase). So far I have...

02 August 2019 1,650 3 View

Is it possible to elute formaldehyde-crosslinked FLAG-tagged protein-RNA complexes from FLAG resins using high salt buffers?

I routinely purify FLAG-tagged protein-RNA complexes that have been crosslinked with formaldehyde using FLAG peptide. Alternatively, it is possible to elute proteins from FLAG resins using high...

04 March 2019 6,712 3 View

Screenings for genes involved in biofilm formation: any suggestions?

In our lab we investigate the biofilm formation of uropathogenic E. coli and I would like to perform a random mutagenesis experiment to be able to identify genes mediating this process. However,...

16 November 2017 7,833 2 View

How to reduce protein aggregation during protein concentration?

I am trying to concentrate the secretome of E. coli using Vivaspin columns (GE). I succesfully concentrate the proteins but when running SDS-PAGE I observe pretty blurry bands and irregular...

21 June 2016 545 9 View

Can membrane dye FM 4-64 affect GFP-tagged protein localization?

I observe dramatic differences in E. coli GFP-tagged protein localization when I analyze non-stained cells and cells stained with membrane dye FM 4-64 GFP-tagged proteins show no specific...

15 May 2015 3,232 6 View

Sonication vs French press for protein localization studies in E. coli: Which is the better option?

I plan to perform cell fractionation and subsequent protein localization in E. coli but I want to disrupt my cells gently so that the protein complexes remain as stable as possible. Sonication is...

25 April 2014 4,530 2 View