S-Nitrosylation-Specific Antibody Introduction

S-nitrosylation is a typical redox-dependent post-translational modification of proteins that plays a crucial role in microrelief regulation within the body and is implicated in numerous diseases. Despite being discovered over 150 years ago, the function of S-nitrosylation in NO-mediated protein modifications has only been studied for the past 20 years. Consequently, a comprehensive investigation and validation of the function of S-nitrosylation are still required. Creative Biolabs offers a wide range of services for the development of S-nitrosylation-specific antibodies utilizing our hybridoma technology platform and phage display platform.

S-Nitrosylation Modification

Nitrosation refers to the covalent incorporation of the nitroso group of NO into another molecule. When nitrosylation occurs at the thiol group of cysteine, it is known as S-nitrosylation. This non-enzymatic and reversible modification process primarily depends on the proximity of the protein to diffusible nitric oxide. S-nitrosylation is commonly observed in mammalian, plant, and even microbial proteins. It acts as a signaling molecule, regulating signaling pathways, and also serves as a mediator for other post-translational modifications to regulate protein function.

S-nitrosylation has been found to target various proteins, including enzymes, receptors, ion channels, and transcription factors. It is closely related to apoptosis, showing a bidirectional effect depending on the cell type, intracellular redox status, and NO concentration. The S-nitrosylation of certain enzymes can influence the course and direction of apoptosis. For instance, the GAPDH enrichment is considered a marker of stimulus-induced apoptosis. Caspases are believed to be involved in the initiation of apoptosis or in the execution of apoptosis. Furthermore, S-nitrosylation can activate matrix metalloproteinase-9, inducing pro-apoptotic effects. In blood-related diseases, S-nitrosylation regulates changes in blood flow in response to external stimuli. The blood oxygen delivery system is a major pathway through which S-nitrosylation affects disease. In diabetes, glycosylation increases hemoglobin's affinity for O2, consequently impacting the release and delivery of S-nitrosylation, thereby promoting diabetic microvascular complications. In neurodegenerative diseases, S-nitrosylation is impacted in the regulation of lipid metabolism, as well as the activity of proteins like Cdk5 and Drp1.

S-Nitrosylation-Specific Antibody

Detecting S-nitrosylation through experimental means is challenging due to the weak S-N bond of S-nitrosylation and its susceptibility to reducing substances and metalloenzymes present in vivo. Currently, two main methods are commonly used for S-nitrosylation detection: the biotin conversion method and the chemiluminescence method. However,both methods have obvious drawbacks. For example, the experimental procedure of the biotin conversion method may damage the fragile S-N bond, and the presence of ascorbate can result in false-positive test results by not only reducing the cysteine sulfhydryl group in the SNO site but also interfering with the interaction with other cysteines. An alternative approach is using antibodies to detect S-nitrosylation, which is equally valid route. However, like the development of all PTM specific antibodies, the development of S-nitrosylation-specific antibodies is not a straightforward process. Since S-nitrosylation is reversible, the protein extraction process should be performed as quickly as possible, kept on ice throughout, and strictly following the provided instructions. In addition, for the isolation and purification of antibodies, they can be purified by binding small peptides corresponding to S-nitrosocysteine to bovine serum albumin (BSA) or keyhole Limpet hemocyanin (KLH), followed by ammonium sulfate precipitation or affinity separation.

Creative Biolabs has a wealth of knowledge and experience about PTM specific antibody discovery. We would be delighted to discuss our knowledge and experience in the development of S-nitrosylation specific antibodies with you.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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