Kinase Profiling
Protein kinases are key regulators of important cellular processes, including growth, stress response, differentiation and apoptosis. About 550 kinases have been identified in the human genome, exerting their functions inside the cell in complex networks. For drug development , signal transduction research and clinical research in-depth insight in the effects of stimuli on multiple protein kinases is often required in order to fully understand the effects of kinase inhibitors, disease development or growth factor stimuli. The PepChip Kinase Profiling products provide this broad view on kinase changes by analyzing kinase activities in cells and tissues on up to 1,184 unique substrates on a single microarray. The low sample volume required allows analysis of biopsies, sections, and white blood cell samples, as well as cultured cells and tumor material.
Cell and tissue lysates can be analyzed on two types of PepChip Kinase substrate microarrays. The PepChip Kinase array contains 1,184 peptides in duplicate. This array is essentially a large substrate collection, covering many peptide substrates found since the early days of kinase analysis. The PepChip Kinomics array contains 1,024 peptides in triplicate derived from known phosphorylation sites from human protein sequences, annotated with reported upstream kinases. In addition, the Pepchip Evaluation slide is used for optimization of the reaction conditions.
PepChip arrays are incubated with cell or tissue lysates. The assay is based on isotopic detection using 33P-labelled ATP. Assay conditions can be adapted, covering a wide range of ATP concentrations, bivalent ion concentration and buffer. Isotopic detection is both highly sensitive and unbiased towards specific phosphorylation sites. Radiolabelled PepChip arrays are imaged, quantified and analyzed to produce a list of substrates undergoing significant phosphorylation changes.
For Full Service Kinase Profiling, cell or tissue lysates are shipped to Pepscan presto on dry ice. Typically, 20-100 micrograms of total protein lysate in 10-50 microliters is required for one array. Biological replicates are recommended. Results are delivered via email, including a summary report, raw data, slide scans and analysis files. Pepchip Kinase arrays are also available as consumables, allowing experiments to be carried out at our customer’s laboratories. Products are supported by the experts from PepScan Presto.
| Product | PC Kinase Array | PC Kinomics Array | PC Kinase Evaluation Slide |
|---|---|---|---|
| Art.no | PCKF00020 | PCKINOM01 | PCKT00010 |
| Substrates | Mixed species (bovine, mouse, rat, pig, human, … etc) |
Human sequences | Mixed species |
| Peptides | 1184 duplicate | 1024 triplicate | 192 duplicate |
| Quantification | PepChip Kinase Grid.zip | PepChip Kinomics Grid.zip | PepChip Trial Grid.zip |
| Substrate-list | PepChip Kinase Map.zip | PepChip Kinomics Map.zip | PepChip Trial Map.zip |
Download Handling manual PepChip: Protocol PepChip for Lysates.zip
Download Manual alignment grids: How to use alignment grids.zip
Protease Profiling
The PepChip Protease array displays as many as 1,024 individual biotinilated protease substrates in triplicate on a glass slide. The specific microarray allows investigation of protease substrate preference and identification of substrates. Uncleaved substrates are detected before and after digestion using commercial available dyes e.g. fluorescein-labeled streptavidin. PepChip Protease arrays are available as consumable and can be used in common arraydetectors.
| Product | PC Protease |
|---|---|
| Art.no | PCPROT01 |
| Substrates | Random sequences |
| Peptides | 1024 triplicate |
| Quantification | PepChip Protease Grid.zip |
| Substrate-list | PepChip Protease Map.zip |
Download Handling manual PepChip: Protocol PepChip for Lysates.zip
Download Manual alignment grids: How to use alignment grids.zip
Custom Design Arrays
Pepscan Presto uses state of the art solid-phase resin based FMOC chemistry for the synthesis of custom peptides in quantities ranging from milligram to gram scale. The peptides are synthesized using modern synthesizers with controlled conditions during synthesis and optimized pre-activation chemistry and double couplings to ensure optimal reaction conditions for the highest quality of peptides. Depending on your requirements, the peptides can be modified at any position in the sequence with labels or fluorescent groups, or labeled or capped at the C- and N-terminus.
Thousands of pre-synthesized peptides designed by the customer can be arrayed in triplicate on multiple glass slides. Designing of the peptides and products are supported by the experts from PepScan Presto.