Identification of Phosphoproteins in Tissues
INQUIRYCreative BioMart provides high-quality customized services and advanced techniques to help our customers identify phosphoprotein in tissues and provide insights into tissue-specific cell signaling. Our scientific team offers their skills, knowledge, and experience to help our customers advance their projects successfully.
Background
Phosphorylation is a common post-translational modification (PTM) that underlies the functional modulation of many proteins in diverse biological processes. Phosphorylation is accomplished through the activity of kinases and can be reversed by phosphatases in a highly dynamic manner. Phosphorylation can alter protein activity and regulate protein-protein interactions, thereby affecting virtually all cellular processes such as cell cycle, signal transduction, and metabolic maintenance. Importantly, deregulation of protein phosphorylation results in a wide range of pathological conditions. Since phosphorylation plays such a role of fundamental importance, it becomes critical to detect protein phosphorylation and assess its functional consequences.
Although many phosphorylation networks have been elucidated using in vitro systems and have provided fundamental information for modeling signal transduction, however, such models cannot be fully applied to ex vivo tissue models and in vivo systems. Moreover, the targeted methods typically used in in vitro cell culture studies would preclude an exhaustive analysis of phosphorylation. Therefore, new methods are in need to be developed for de novo identification and characterization of phosphoproteins from tissues. Analytical datasets of phosphorylation studies collected using human tissue specimens will offer great promise for elucidating pathogenic protein signaling networks and developing targeted therapies.
Figure 1. Cross-Tissue Comparison of Protein and Phosphoprotein Expression. (Huttlin E L, et al., 2010)
Service Offering
Creative BioMart offers a comprehensive service offering for the identification of phosphoproteins in different types of plant and animal tissues. We can handle different tissue sample collection formats including fresh frozen tissues, formalin-fixed paraffin-embedded tissues, and tissue extracts. Our scientists are dedicated to providing our customers with end-to-end services to help them save time and effort.
Detection and Quantitation of Phosphoproteins in Tissues
The development of phospho-specific antibodies has made the detection of phosphoproteins possible. We offer diverse techniques, including immunohistochemistry, immunofluorescence, and Western blot, for the detection of phosphoproteins in tissue samples. We also provide optimized quantum dot-based immunofluorescence technique for in situ quantitation of phosphoproteins.
Phosphoprotein Enrichment from Tissue Samples
The low abundance of phosphoproteins adds to the difficulty of their study, which makes it crucial to develop and optimize protocols for phosphoprotein enrichment. We have optimized a number of techniques to preferentially enrich phosphoproteins from tissue samples, including but not limited to immobilized metal affinity chromatography (IMAC), immunoprecipitation (IP), metal-oxide affinity chromatography (MOAC), and enrichment by chemical modification.
Phosphoprotein Identification
Identification of the phosphorylation status of proteins in tissues has become an indispensable method for unraveling signaling networks. We have established an advanced mass spectrometry (MS) platform to provide informative data for phosphoprotein identification. Our scientists can also perform functional annotation of identified phosphoproteins.
Creative BioMart is an advanced biotech company within the field of kinase/phosphatase biology. We offer our expertise in establishing assays to analyze tissue-specific phosphorylation. The extensive experience of our scientific team translates into the robust wealth of knowledge that we use to provide you with the highest quality of service. If you are interested in any of our services or have additional questions, please do not hesitate to contact us.
References
- Huttlin E L, et al. A tissue-specific atlas of mouse protein phosphorylation and expression. Cell, 2010, 143(7): 1174-1189.
- Garaguso I & Borlak J. A Rapid Screening Assay to Search for Phosphorylated Proteins in Tissue Extracts. PloS one, 2012, 7(11): e50025.