Applications/Post-translational modifications

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Setting the standard for protein and peptide applications
Proteomics/Biomarker Discovery
SELECT
Characterization of Biopharmaceuticals
SELECT
Targeted analysis
SELECT
Post-translational modifications

Post-translational modifications

THIS IS HOLDING TEXT Classic discovery proteomics is most commonly carried out using data-dependent acquisition (DDA) strategies. While it is easy to setup data acquisition and analysis to obtain a high proteome depth, the reproducibility is commonly low due to stochastic sampling. In contrast, data-independent acquisition (DIA) allows for more accurate and sensitive protein quantification and has become widely used. Our iST kits are suitable for experiments with metabolic labeling or in label-free mode, whereas our iST-NHS kits are suitable for experiments with chemical labeling such as iTRAQ or TMT.

Cell pellet

Add  50 µL of our LYSE or LYSE-NHS buffer directly to the cell pellet and proceed with regular protocols

FACS-sorted cells:

If the volume   of FACS sorted cells exceeds 11 µL, centrifuge  to take off the supernatant and decrease the volume before adding lysis buffer. Alternatively use the higher concentration lysis buffers.

Cell line lysates:

Certain lysis buffers are compatible with our iST and iST-NHS workflows. For further details please see our FAQ 1.1 If they are not compatible protein precipitation may be required.

Adherent cell culture samples

If extracellular matrix or transmembrane proteins are not of interest, you can use cell culture-grade trypsin to detach the adherent cells, wash them once with PBS and then store the cell pellet fraction as input for our regular protocols. Alternatively, if extracellular matrix or transmembrane are of interest and affected by partial trypsin digestion, add 50-100 µL of our LYSE / LYSE-NHS buffer directly to the cells and scrape them off using a rubber policeman. Collect the scraped material and heat to 95°C for 10 min before continuing with the regular iST / iST-NHS protocol.

Protocols:
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PreOmics Values

Smile

Improved sample quality

High reproducibility: R
2
>0.9
Working range: 1-100 µg
Increased peptide and protein identifications

Relax

Quicker methodology

Less than 2.5 hours hands-on time
Single step incorporating lysis, reduction and alkylation
All-in-one solution, all reagents provided
Compatible with standard lab equipment

Realise

Cleaner peptides ready for analysis

Dual clean up step removes hydrophilic and hydrophobic contaminants
Achieve ready-to-measure peptides
Reduce mass spectrometry downtime
Manual to fully automated processing options available

Imagine

Applying the science
Find out more about the wide range of applications and sample
types to which PreOmics technology has been successfully applied