Applications/Sample types/Cell lines

Plan! Use PreOmics' products for your research

Resources for working with cell lines
Cell lines
Model Organisms
SELECT
Tissue/FFPE
SELECT
Plant
SELECT
Biological Fluids
SELECT
Immunoprecipitation
SELECT

Cell Lines

Cell lines and primary cells can be directly processed with PreOmics sample preparation technology for a wide range of applications. Typical experiments may include protein-binding interactions, pathway elucidation, bottom-up proteomics and semi-targeted work. For example when testing cell therapies assessing the up-regulation or down-regulation of the target protein is essential.

The iST/iST-NHS kits are suitable for use in research and regulated environments. The kits contain all you need for your peptide or protein sample preparation and are suitable for use by all members of your lab, no specialist experience is needed. The flexible formats of single samples or racked in to 96 well holders allow the kits to be used manually or with a range of automation options. The cartridges can be processed by centrifugation or using positive pressure manifolds, again giving options for all labs. Dedicated resources are shown below and our technical support team would be happy to help with advice.

Please always ensure the total protein present is within the 1 – 100 µg range. In our labs we have established that working to a set OD or cell count is more reproducible than a total protein concentration assay, initially you may need to make measurements of total protein vs OD or cell count until you have a good understanding of the relationship. Guidelines are given below dependent on the starting state of the cell line material. Cell lines and primary cells can be directly processed with PreOmics sample preparation technology for a wide range of applications. Typical experiments may include protein-binding interactions, pathway elucidation, bottom-up proteomics and semi-targeted work. For example when testing cell therapies assessing the up-regulation or down-regulation of the target protein is essential.

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.

PreOmics Values

Smile

Improved sample quality

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

Relax

Quicker methodology

Less than 2.5 hours total 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