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Questions related from Mikel Irastortza
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...
08 August 2019 8,002 0 View
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...
03 March 2019 7,022 3 View
Due to short sequence length and strong secondary structure I find difficult to target CRISPR gRNAs by qPCR. The sequence of the gRNA I am targeting is...
04 April 2018 6,893 1 View
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,...
11 November 2017 8,570 3 View
LB is not compatible with formaldehyde in culture cross-linking of E. coli because amine groups derived from yeast extract and peptone interact with the cross-linking agent. Is there any...
06 June 2017 7,187 4 View
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...
06 June 2016 1,682 10 View
I routinely perform cell fractionation and subsequent protein localization studies with Escherichia coli. All protocols I have come across used EDTA as chelating agent to avoid proteolysis during...
06 June 2014 6,213 6 View
I have been searching for commercial antibodies against E. coli membrane proteins but I could not find any. Could anybody, please, let me know about some provider offering these antibodies? In...
06 June 2013 3,244 4 View
I am trying to purify membrane protein complexes in E. coli. Briefly, I perform high speed (100000 x g) ultracentrifugation to separate the cytosolic fraction and then wash the membrane fraction...
06 June 2013 2,367 24 View
