DUOLINK IMAGE TOOL SOFTWARE

The dedicated Duolink ImageTool software available from Olink makes it easy to automatically and accurately quantify the signal levels for each experimental condition either as an average number of signals per cell in each region or image of interest or as individual cell data, even for users unfamiliar with more complex image analysis suites. The interaction was only detected in the membrane of oocytes and not in the junction between two oocytes during the two-cell stage.Įasy analysis and quantification-retrieve single-cell data A recent report describes how Duolink is used to visualize the previously undetected interaction between fibroblast growth factor 2 and neuronal cell adhesion molecule in single-cell oocytes 4. Every signal is physically linked to its target and stays in the correct cellular locale as the assayed interaction takes place. The strong and highly specific amplification of individual, detected protein interaction events ensures that every signal is easily visible as a distinct spot in a fluorescence or brightfield microscope. Visualization of protein interactions-see what's really there This highlights the importance of being able to study even transient and weak interactions to fully understand complex biological systems. In a recent report, researchers studied VEGF receptor dimerization patterns, and although VEGF-C ligand–induced heterodimerization between VEGFR2 and VEGFR3 was detected using both Duolink and Co-IP, VEGF-A ligand–induced heterodimerization was only found using Duolink 3. The fixation 'freezes' the cells at a desired time point and allows one to use a time-series of fixation to obtain precise measurement of the dynamics of a cellular process that has been perturbed. Protein–protein interactions play a pivotal role in normal cellular functions as well as in carcinogenesis.Using Duolink directly answers the crucial question of whether the interaction takes place in the cell when all the necessary proteins are present in their physiological context and at physiologically relevant concentrations.ĭuolink detects interactions you cannot detect with Co-IPĭuolink utilizes fixed cells or tissue, ensuring that even transient complexes are intact at the time of measurement. The protein–protein interactions form functional clusters during signal transduction. To elucidate the fine calibration of the protein–protein interactions of protein tyrosine phosphatase interacting protein 51 (PTPIP51) a small molecule drug, namely LDC-3, directly targeting PTPIP51 is now available. Therefore, LDC-3 allows for the studying of the regulation of the endogenous interactome by modulating PTPIP51 binding capacity. Small interfering ribonucleic acid (siRNA) experiments show that the modification in PTPIP51 binding capacity is induced by LDC-3. Application of LDC-3 annuls the known regulatory phosphorylation mechanisms for PTPIP51 and consequently, significantly alters the assembly of the PTPIP51 associated protein complexes.

The treatment of human keratinocytes (HaCaT cells) with LDC-3 induces an altered protein–protein interaction profile of the endogenous interactome of PTPIP51.

In addition, LDC-3 stabilizes PTPIP51 within a mitogen activated protein kinase (MAPK) complex composed of Raf-1 and the scaffold protein 14-3-3, independent of the phosphorylation status of PTPIP51.

Of note, under LDC-3 treatment the regulatory function of the PTP1B on PTPIP51 fails to impact the PTPIP51 interaction characteristics, as reported for the HaCaT cell line. In summary, LDC-3 gives the unique opportunity to directly modulate PTPIP51 in malignant cells, thus targeting potential dysregulated signal transduction pathways such as the MAPK cascade. The provided data give critical insights in the therapeutic potential of PTPIP51 protein interactions and thus are basic for possible targeted therapy regimens. Protein tyrosine phosphatase interacting protein 51 (PTPIP51) was identified in various protein complexes fulfilling opposite cellular functions, namely proliferation, differentiation and apoptosis. Meanwhile, a growing number of PTPIP51 interaction partners have been described.

Duolink image tool