S-Nitrosylation Analysis Service

Introduction

S-Nitrosylation refers to a covalent attachment of an NO moiety to sulfhydryl residues of proteins. The sulfhydryl residues belongs to a subset of specific cysteine residues in proteins, the resulting SNO is an S-nitrosoprotein. SNOs have a short half-life in the cytoplasm because of the host of reducing enzymes, including glutathione (GSH) and thioredoxin, that denitrosylate proteins. Therefore, SNOs are often stored in membranes, vesicles, the interstitial space and lipophilic protein folds to protect them from denitrosylation. For example, caspases, which mediate apoptosis, are stored in the mitochondrial intermembrane space as SNOs.

Similar to phosphorylation, S-Nitrosylation is a reversible precess. The denitrosylation is an enzymatic catalyzing process that reverses the S-Nitrosylation process. However, S-nitrosylation is not a random event, and only specific cysteine residues are S-nitrosylated. Under physiologic conditions, protein S>-nitrosylation and SNOs provide protection preventing further cellular oxidative and nitrosative stress. Aberrant S-Nitrosylation may lead to protein misfolding, synaptic damage, and apoptosis. Dysfunction of the SNO signaling has been implicated in the pathogenesis of many diseases, such as Alzheimer’s disease cardiovascular diseases.

S-Nitrosylation

Our S-nitrosylated Proteins Analysis Service

Because proteins may contain multiple cysteines and due to the labile nature of SNOs, S-nitrosylated cysteines can be difficult to detect and distinguish from non-S-nitrosylated amino acids. Creative Proteomics has established a highly sensitive HPLC-MS/MS pipeline that can analyze S-nitrosylated cysteines in both eukaryotic and prokaryotic organisms. In addition, we have optimized our protocol, to enable more fast and sensitive site mapping service for S-nitrosylated cysteines.

We rely on Thermo Fisher's Orbitrap Fusion Lumos mass spectrometry platform combined with nanoLC-MS/MS to provide S-nitrosylated Proteins analysis service packages. We will take care of all aspects of the project, including protein extraction, proteolytic cleavage, acylated peptide enrichment, peptide separation, mass spectrometry analysis, raw mass spectrometry data analysis, and bioinformatics analysis.

Workflow of our S-Nitrosylation analysis service:

Workflow of our S-Nitrosylation analysis service

In gel or in solution digestion of proteins
Enrichment of S-nitrosocysteine using anti-S-nitrosocysteine antibody
HPLC separation, followed by ESI-TOF MS/MS analysis
Mass spectrometry data interpretation

Technology platform:

Advantages

Applicable to a variety of processing methods: direct mass spectrometry detection of proteins, or detection after conversion to Biotin, His or other tags.

High precision and accuracy: Using a high-resolution Thermo Fisher mass spectrometer, specific modification sites and peptides can be detected.

Simple, fast and convenient: The operation is simple and convenient; the detection and experiment period is short, and the results are reliable.

Ordering Procedure:

Ordering Procedure

Proteomics Sample Submission Guidelines

Ensure your samples are prepared and submitted correctly by downloading our comprehensive Proteomics Sample Submission Guidelines. This document provides detailed instructions and essential information to facilitate a smooth submission process. Click the link below to access the PDF and ensure your submission meets all necessary criteria.

Download

* For Research Use Only. Not for use in diagnostic procedures.

Our customer service representatives are available 24 hours a day, 7 days a week. Inquiry

From Our Clients

"I recently used their proteomics service for a project analyzing protein interactions in yeast models. The team was very responsive and helped clarify the methodology they employed, which made me feel confident in the results. The data quality was solid, with clear identification of several key proteins involved in our study. Their thorough analysis enabled me to pinpoint specific interactions that I hadn't considered before, which significantly improved the direction of my research. I appreciate their professionalism and support throughout the process."

Sarah Thompson, University of California, Berkeley

"Our lab collaborated with them on a project studying cancer biomarkers. The proteomics analysis provided was detailed and focused, specifically highlighting the differential expression of proteins between healthy and tumor samples. Their clear explanations of the data helped my team understand the biological implications. I also appreciated their willingness to revise the reports based on our feedback, ensuring that we had everything we needed for our publication. This collaborative spirit was invaluable."

Emily Rodriguez, Stanford University

"Our lab worked with them on a project studying the effects of diet on gut microbiota using proteomics. They used a label-free quantification method to analyze proteins in fecal samples before and after dietary intervention. The results showed significant changes in protein expression linked to microbial activity. This was pivotal for our hypothesis about diet-microbiota interactions. The clarity of their data presentation made it easy for our team to integrate these findings into our ongoing research."

Dr. Lisa Wong, University of Toronto

"My experience with Creative Proteomics during the mass spectrometry analysis was excellent. We sent in human saliva and mouse brain tissue samples, which they expertly analyzed using both LC-MS and GC-MS techniques. The results were invaluable, revealing key metabolites in the saliva and identifying biomarkers linked to brain function in the brain tissue."

Dr. Emily Carter, Senior Research Scientist

"The overall service from Creative Proteomics was outstanding. They made the entire process seamless and efficient, allowing us to focus on our research. We worked with leaf and root samples from various Arabidopsis genotypes for targeted metabolomics analysis. Their thorough profiling of primary and secondary metabolites gave us important insights into how the plants respond metabolically to environmental stress."

Dr. Laura Henderson, Plant Physiologist

"We had a pleasant collaboration with Creative Proteomics on mass spectrometry analysis of lipids. They conducted a detailed analysis of lipid species, providing us with important insights into lipid metabolism and its relationship with metabolic syndrome disease states."

Dr. Sarah Mitchell, Research Scientist

Online Inquiry

Please submit a detailed description of your project. We will provide you with a customized project plan to meet your research requests. You can also send emails directly to for inquiries.

Great Minds Choose Creative Proteomics