NGB DIAGNOSTICS PRIVATE LIMITED

GLOBAL PROTEOME PROFILING

Global proteomic analysis and profiling assays allow you to identify and accurately measure hundreds to thousands of proteins in nearly any sample type which contains protein.

Standard Label-Free Quantitative Proteome Profiling Assays

These assays are appropriate for a wide range of samples types :

1 . Affinity-Enriched Proteome Profiling

ID and Quantify : Up to 500 protein groups (redundant isoforms do not count to this total)
Specifications : UPLC-MS/MS, 25 cm column, up to 45,000 sequencing events
Suggested sample types : Affinity-enriched and other minimally complex samples

2 . Prokaryotic Proteome Profiling

Suggested sample types : Prokaryotes, some eukaryotes (yeast), some biofluids (saliva, urine)
ID and Quantify : Up to 4,000 protein groups (redundant isoforms do not count to this total)
Specifications : UPLC-MS/MS, 25 cm column, up to 45,000 sequencing events
Assay : LFQ Profiling, Top 4,000

3 . Plasma / Serum Label-Free Quantitative Proteome Profiling Assays

ID and Quantify : Up to 500 protein groups (redundant isoforms do not count to this total)
Specifications : UPLC-MS/MS, 25 cm column, up to 45,000 sequencing events
Assay : Non-depleted Plasma / Serum Proteome Profiling
Sample : At least 5 uL of plasma or serum

Sample Requirements

See each assay for assay-specific requirements, in general samples should :

contain at least 10 micrograms of total protein, 30 micrograms is ideal
be free of contaminants such as detergents or protease inhibitors, please let us know if this is not the case
ideally be less than 100 uL in volume, but no greater than 1.5 mL, a handling charge applies to larger volumes

Workflow

Enzymatic digestion with sequencing-grade trypsin
Protein extraction and cleanup
SPE of the digest
UPLC-MS/MS
- UPLC - Thermo Easy-nLC 1200 fitted with a heated Easy-Spray column of 25 or 50 cm length
- MS/MS - Thermo Q-Exactive HF-X quadrupole-Orbitrap mass spectrometer
Peptide identification with Comet and Tandem, spectral library searching and match-between-runs
Data securely stored and accessible for one year with our basic plan
Quantitation via MS1 peak are measurement

Deliverables

All data is hosted and delivered using our web-based data portal
View data reports online or download as Excel files
Data reports include:
- proteins grouped by common peptide identifications, to allow isoform discrimination
- tandem MS spectrum viewer: see fragment ion assignments for all identified peptides
- proteins identified, with intensity values for each sample
Download raw data files (mzML format)

Optional deliverables available at extra cost include a growing number of add-on reports :

Protein LFQ Comparison: fold-change and statistical significance measurements for up to 5 comparisons between any samples or groups of samples in the project
Taxonomy Classification: lowest-common ancestry calculation of discriminative peptides in a sample

GLOBAL PROTEOME PROFILING

Standard Label-Free Quantitative Proteome Profiling Assays

Sample Requirements

Workflow

Deliverables

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