6,041 Result(s)
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Protocol
Methods for Intracellular Peptidomic Analysis
Peptides have broad biological significance among different species. Intracellular peptides are considered a particular class of bioactive peptides, whose generation is initiated by proteasomal degradation of ...
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Protocol
Quantitative Peptidomics Using Reductive Methylation of Amines
A number of different approaches have been used for quantitative peptidomics. In this protocol, we describe the method in which peptides are reacted with formaldehyde and sodium cyanoborohydride, which convert...
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Protocol
Quantitation of MUC5AC and MUC5B by Stable Isotope Labeling Mass Spectrometry
Mucins MUC5AC and MUC5B are large glycoproteins that play an essential role in the innate defense of epithelial surfaces and their quantitation in biological samples would be informative about the health statu...
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Protocol
Nontargeted Identification of d-Amino Acid-Containing Peptides Through Enzymatic Screening, Chiral Amino Acid Analysis, and LC-MS
d-amino acid-containing peptides (DAACPs) in animals are a class of bioactive molecules formed via the posttranslational modification of peptides consisting of all-l-amino acid residues. Amino acid residue isomer...
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Protocol
Bioinformatics for Prohormone and Neuropeptide Discovery
Neuropeptides and peptide hormones are signaling molecules produced via complex posttranslational modifications of precursor proteins known as prohormones. Neuropeptides activate specific receptors and are ass...
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Protocol
Eliminative Oximation of O-Glycans from Mucins
The large variety and high concentration of O-glycans are characteristic properties of mucins and play a crucial role in their unique functions. Analyzing the O-glycans of mucins is essential for investigating th...
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Protocol
An Updated Guide to the Identification, Quantitation, and Imaging of the Crustacean Neuropeptidome
Crustaceans serve as a useful, simplified model for studying peptides and neuromodulation, as they contain numerous neuropeptide homologs to mammals and enable electrophysiological studies at the single-cell a...
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Protocol
Liquid Chromatography and Capillary Electrophoresis Analysis of 2AA-Labeled O-Glycans
To reveal O-glycan structures in mucins, it is necessary to release covalently bound O-glycans from the polypeptide backbone and derivatize to a form suitable for structural analysis. Various derivatization metho...
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Protocol
Peptidomics of Zebrafish Brain in a 6-OHDA-Induced Neurodegeneration Model
Bioactive peptides such as neuropeptides and peptide hormones are largely understood in their involvement in a variety of physiologic systems. In addition to the neuropeptides produced and processed by the cla...
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Protocol
Intracellular and Extracellular Peptidomes of the Model Plant, Physcomitrium patens
Here, we report our approach to peptidomic analysis of the plant model Physcomitrium patens. Intracellular and extracellular peptides were extracted under conditions preventing proteolytic digestion by endogenous...
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Protocol
MALDI-TOF MS/MS Analysis of Permethylated O-Glycan Alditols Released from Mucins
Mucin glycans are associated with the function of mucin in maintaining mucosal homeostasis. Therefore, the glycomic analysis of mucins is crucial. Matrix-assisted laser desorption/ionization time-of-flight mas...
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Protocol
Assessment of Mucin-Associated Gene Expression Levels on the Ocular Surface
The ocular surface is covered with a mucus layer. The mucin-associated genes expressed in the ocular surface cells include MUC1, MUC4, MUC5AC, and MUC16. Impression cytology is useful for collecting specimens fro...
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Protocol
Identification of Peptides in Spider Venom Using Mass Spectrometry
Spider venoms are composed of hundreds of proteins and peptides. Several of these venom toxins are cysteine-rich peptides in the mass range of 3–9 kDa. Small peptides (<3 kDa) can be fully characterized by mas...
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Protocol
Peptidomics Strategies to Evaluate Cancer Diagnosis, Prognosis, and Treatment
Peptides have potential bioactive functions, and the peptidomics landscape has been broadly investigated for various diseases, including cancer. In this chapter, we reviewed the past four years of literature a...
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Protocol
Differential Glycoform Analysis of MUC1 Derived from Biological Specimens Using an Antibody-Overlay Lectin Microarray
The association between altered glycosylation of MUC1 and various disease events has sparked significant interest. However, analytical technologies to investigate the disease-related glycoforms of endogenous M...
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Protocol
Expression Analysis of Genes Corresponding to Mucins and Their Glycans from Cervical Tissue Using RNA Sequencing
In the female reproductive tract, mucin proteins are the main component of mucus secreted by cervical goblet cells and play an essential role in many biological functions. They act as a medium for lubrication ...
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Protocol
Extraction of Mucin from Rodent Feces and Determination of O-Linked Oligosaccharide Chain Equivalent Derived from Fecal Mucin
Since the core protein of mucin in the digesta of the stomach and small intestine, which is less affected by bacteria, remains intact, mucin content can be measured by enzyme-linked immunosorbent assay (ELISA)...
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Protocol
Profiling Human Cerebrospinal Fluid (CSF) Endogenous Peptidome in Alzheimer’s Disease
Human cerebrospinal fluid (CSF) is a rich source for central nervous (CNS)-related disease biomarker discovery due to its direct interchange with the extracellular fluid of the CNS. Though extensive proteome...
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Protocol
Analysis of the Interaction Between Mucin and Green Fluorescent Protein (GFP)-Tagged Galectin-2 Using a 96-Well Plate
Due to a significant proportion of glycans binding to the peptide (constituting approximately 50–90% of the molecular weight), analyzing the interaction between the entire mucin molecule and its recognition pr...
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Protocol
Isolation of Membrane Bound Mucins from Human Bronchial Epithelial Cells
Membrane-bound mucins constitute a large portion of the periciliary layer of lung epithelial surfaces, and thus play an important role in many aspects of innate defense. The biophysical and biochemical propert...
