Michael Plank

10 Questions 21 Answers 0 Followers

Questions related from Michael Plank

What should be distance between promoter and initiator-ATG?

Hi, I was wondering if anyone here has some insight into what should be the distance between the promoter and the translation start site / initiator-ATG when making a DNA construct? In...

03 October 2021 8,384 2 View

Does anyone have a protocol for denaturing IP from S. cerevisiae?

Hi, would anyone have a good protocol for denaturing IP from S. cerevisiae? I want to look at the phosphorylation state of a protein and therefore lyse the cells under harsh conditions that...

11 May 2021 3,927 0 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

02 March 2021 748 1 View

Do you have recommendations for plasmids for yeast protein tagging?

Hi, would anyone have recommendations for a set of plasmids for endogenous C-terminal protein tagging in S. cerevisiae (i.e. plasmids as template for the PCR product to transform). I`m looking for...

19 November 2020 4,054 1 View

Formylation artefacts in proteomics sample preparation?

Hi all, I was analysing a phospho-proteomics sample (tryptic digest from a human cell line) and, as the number of phosphosites I could identify was surprisingly low, I ran MaxQuant with dependent...

29 April 2019 2,818 1 View

How to perform on-filter on-bead digestion?

Hi all, I`m trying to analyse PTMs (mainly phosphorylation) of a yeast protein by MS after enrichment of the tagged (e.g. TAP-tag) protein. I would like to perform the tryptic digestion on the...

02 October 2018 9,724 6 View

Protocol/settings for yeast proliferation assay in 96-well plates?

Hi, would anyone have a protocol or be willing to share their settings for a yeast (S. cerevisiae) proliferation assay in 96-well plates? We have a Tecan Spark 10M platereader, but I`d imagine...

20 June 2018 2,898 10 View

Does anyone know what peak of m/z 524.739 (charge 2+) in LC-MS could be?

Hi all, I`m frequently observing this peak at the very beginning of the elution of my nanoLC-MS/MS samples (mainly phosphoproteomics). It`s pretty abundant and elutes on a much wider RT range than...

02 March 2017 3,160 6 View

How do I best SILAC lable (non-auxotrophic) yeast? Starvation prior to labelling?

Hi, when I try to SILAC lable a yeast strain which is not arginine auxotrophic with heavy arginine I generally gel low labelling efficiencies. In mammalian cell culture people often starve their...

23 September 2015 2,851 1 View

How can I use information from different protease digests for PTM localization?

Hi, it should be possible to enhance your confidence in PTM site localization by combining information from digests with different proteases, e.g. by considering where the peptides overlap and...

02 March 2015 3,792 3 View