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PhosphoDetect™ Phospho-specific Antibodies
 | | | The Importance of Protein Phosphorylation-Dephosphorylation Protein phosphorylation-dephosphorylation is one of the major signaling mechanisms for modulating the functional properties of proteins involved in gene expression, cell adhesion, cell cycle, cell proliferation, and differentiation. Proteins can be phosphorylated on specific serine, threonine, or tyrosine residues by the action of protein kinases. Protein kinases catalyze the transfer of a phosphoryl group from a high-energy compound, such as ATP, to a nucleophilic acceptor group located on the amino acid side-chain(s) of proteins. Protein phosphatases are phosphoesterases that catalyze the hydrolytic removal of a phosphate group from hydroxylated amino acid residues on proteins. In contrast to permanent covalent modifications of proteins, this reversible phosphorylation-dephosphorylation reaction is a fast and efficient way to change the properties of proteins in the compartmentalized cellular environment. In order to sustain target specificity among hundreds of phosphoproteins within a cell, cells maintain a vast network of delicately regulated protein kinases and phosphatases. Due to the fact that many of these protein kinases are implicated in cancer, as well as neurodegenerative diseases, researchers are actively seeking drug candidates that can control the activity of particular protein kinases. The most obvious method for screening a drug candidate for efficacy is to determine its effects on the phosphorylation state of either the protein kinase or its substrate. Until recently, radioactivity-based (32P or 33P) assays were the best method for assessing changes in phosphorylation. Phospho-specific
antibodies have
emerged as important
tools for studying signal
transduction pathways |
Although phosphoproteins account for only 10-20% of the total proteome, their dynamic nature makes them important regulatory targets in the cell. The ability to determine the state of phosphorylation of specific proteins is of great value in the pursuit to establish the function of a given protein. In recent years, phospho-specific antibodies, which can discriminate between the phosphorylated and non-phosphorylated forms of a given protein, have become the tools of choice for analysis of protein phosphorylation. Phospho-specific antibodies are novel tools for qualitative and quantitative detection of phosphorylated proteins without the risks associated with radioactivity. Phospho-specific antibodies are affinity-purified and are usually depleted for cross-reactivity with non-phosphorylated form of proteins, enabling them to detect a specific protein in a complex mixture of proteins in cells. They recognize the phosphorylated amino acids in the context of the surrounding amino acid sequence. Hence, using a combination of pan antibody and phospho-specific antibody, you can assess the degree of phosphorylation of any given protein.
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| Why PhosphoDetect™ Phospho-specific Antibodies?PhosphoDetect™phospho-specific antibodies discriminate between phosphorylated and non-phosphorylated forms of proteins or amino acids with exquisite specificity. These phospho-specific antibodies are suitable for many applications, including Western blot analysis, immunofluorescence microscopy, ELISA, immunocytochemical assays, and flow cytometry, thus enabling researchers to study the dynamics of protein phosphorylation in vivo, in situ, and in real time. The PhosphoDetect brand is your guarantee that our phospho-specific antibodies meet the rigid quality standards you rely on for consistent experimental results. PhosphoDetect™ Anti-ATM (pSer1981) Mouse mAb (10H11.E12),
Cat. No. DR1002
Detection of ATM phosphorylated on Ser1981. HeLa cells were treated with 10 mM camptothecin for 2 hrs and fixed with 100% methanol. Cells were probed with PhosphoDetect™ Anti-ATM, (Ser1981) at 1.5 mg/ml. Fluorescently conjugated donkey anti-mouse secondary was used to visualize phosphorylated ATM. Cells were co-stained with DAPI (blue) at 1 mg/ml to visualize nuclei.
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