Effect of flash freezing of pelleted cells?

01 February 2017 1,646 3 View

Can MSCs be grown on a glass coverslip?

I am trying to grow mesenchymal stem cells on a glass coverslip (to be used for confocal microscopy). Does anyone have any experience with growing (not simply adhering) MSCs to glass coverslips?...

10 November 2014 7,159 4 View

Are there any post translational modifications that include addition of a thiol?

I know there are PTMs that make use of the thiol group on cysteine, but does anyone know of any PTMs that add a thiol group to a protein (in any organism, not just humans)?

02 March 2013 1,323 1 View

What controls are needed in immunoprecipitation?

I have a custom antibody (it works for Western blot and immunohistochemistry) that I would like to try for immunoprecipitation. I know that the antibody successfully binds my beads (I'm...

02 March 2013 4,804 7 View

Chemical lysis of coelomocytes?

Does anyone know of a chemical lysis method for total protein isolation from coelomocytes? I'm currently using physical disruption in the presence of detergent. This gets me a fair amount of my...

31 December 2012 4,234 5 View

How can you evaluate EDU (labeling of newly synthesized DNA) in cells that are non adherent without flow cytometry?

We are studying cell proliferation in a cell type (from a marine invertebrate) that is non-adherent. Does anyone have suggestions for method(s) to evaluate EDU labeling in non adherent cells? We...

08 September 2012 3,714 17 View

Good tool for qRTPCR primer design?

I'm designing some primers for qRTPCR. Does anyone have any suggestions for a good qRTPCR primer design tool? (I expect to need to check for hairpins and dimers and what not, but would prefer...

08 September 2012 1,395 5 View

Suggestions for immunoprecipitation kits/protocols?

I'm somewhat new to IP and am looking for a good kit (or bead type) to isolate my proteins (large family of related proteins; we have an antibody specific for a conserved region). My antibody is a...

07 August 2012 5,033 5 View

How to evaluate EDU in cells that are non-adherent without flow cytometry?

We are studying cell proliferation in a cell type (from a marine invertebrate) that is non-adherent. Does anyone have suggestions for method(s) to evaluate EDU labeling in non adherent cells?...

07 August 2012 598 1 View

Has anyone tried using Bio-Rad Criterion XT gels with a home-made running buffer?

I'm looking to buy gels for the second dimension of IEF, and am attracted to the Bio-Rad Criterion XT gels - largely for their relatively long shelf-life (either the Bis-Tris with MOPS running...

07 August 2012 3,491 12 View

Anyone that can help with Maxquant TMT16plex which is missing?

Hi I am very new to proteomics analysis. I have got raw files from core facility running thermofisher TMT 16plex 4 hours. I cannot find on maxquant the option TMT16Plex there is only up to 11...

25 February 2021 1,675 3 View

How to manage western blot for proteomics data validation?

Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...

23 February 2021 6,671 4 View

Unable to detect a truncated protein variant, but I can observe a truncated cDNA from Sanger?

Hi, so basically... I am trying to mimic a clinically observed condition wherein a frame-shift mutation happens only at one allele of the patients, giving rise to a truncated variant which evades...

17 February 2021 1,130 6 View

Is it possible to remove the cells from a tissue and keep only the extracellular matrix for proteomics?

Hello! I would like to know if there is a way to flush/remove all the cells out of a tissue and keep only the extracellular matrix, in order to analyse its composition using e.g. proteomics....

14 February 2021 1,602 4 View

Apart from MOFA what other software is available for analysis of a multi-omic study?

Study bringing together genomics, proteomics etc together

02 February 2021 2,856 2 View

How can I reduce run-to-run variation in TMT MS experiments?

I planned an experimental setup using 10-plex TMT with 9 conditions and 1 bridge channel to compare 5 replicates. But since I experienced some run-to-run variation, I was wondering if a better...

31 January 2021 3,405 3 View

Where can I purchase NanoLC emitter fused silica picoTip?

Hi, I'm working with NanoLC/MS for both Metabolomics and Proteomics. We usually make our nano columns using emitters from NewObjective. Unfortunately New Objective, due to the pandemic, is now...

10 January 2021 1,176 4 View

An optimal protocol of EVs isolation for proteomics?

Hi everyone, I am starting a project in analyzing EVs proteomics in cancers. However, the most published articles I have found stated only brief isolation protocols that could not fully be...

17 December 2020 7,600 2 View

Infected plant protein lysate washing out purified GST-tagged protein on agarose resin?

Hi, I am trying to screen infected plant proteomes for targets of a secreted pathogen protein. I have purified the pathogen protein using a GST-tag on an agarose resin. I am using the Pierce...

23 November 2020 7,009 3 View

Dataset: Orbitrap velos CID MS/MS spectra ?

Hi, I want the MS2-CID spectra from orbitrap velos mass spectrometry of organism: human (Mod: C+57) for testing my peptide identification algorithm developed. Large set of orbitrap velos dataset...

16 November 2020 8,261 1 View