N-glycosidase F
N-glycosidase F - Related Videos
09:10
Synthesis of Indoxyl-glycosides for Detection of Glycosidase Activities
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Cited by 4 •
2015
Indoxyl glycosides are well-established and widely used tools for enzyme screening and enzyme activity monitoring. Especially for glucose type structures previous syntheses proved to be challenging and low yielding. Our novel approach employs indoxylic acid esters as precious intermediates to yield a considerable number of indoxyl glycosides in good yields.
06:11
Unfractionated Bulk Culture of Mouse Skeletal Muscle to Recapitulate Niche and Stem Cell Quiescence
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Cited by 4 •
2023
Skeletal muscle comprises multiple cell types, including resident stem cells, each with a special contribution to muscle homeostasis and regeneration. Here, the 2D culture of muscle stem cells and the muscle cell niche in an ex vivo setting that preserves many of the physiological, in vivo, and environmental characteristics are described.
12:21
Stimulated Stokes and Antistokes Raman Scattering in Microspherical Whispering Gallery Mode Resonators
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Cited by 1 •
2016
Efficient generation of nonlinear phenomena related to third order optical non-linear susceptibility Χ(3) interactions in triply resonant silica microspheres is presented in this paper. The interactions here reported are: Stimulated Raman Scattering (SRS), and four wave mixing processes comprising Stimulated Anti-stokes Raman Scattering (SARS).
09:02
Protocol for the Differentiation of Human Induced Pluripotent Stem Cells into Mixed Cultures of Neurons and Glia for Neurotoxicity Testing
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Cited by 45 •
2017
Human induced pluripotent stem cells (hiPSCs) are considered a powerful tool for drug and chemical screening and for the development of new in vitro models for toxicity testing, including neurotoxicity. Here, a detailed protocol for the differentiation of hiPSCs into neurons and glia is described.
08:37
Immunostaining of Whole-Mount Drosophila Testes for 3D Confocal Analysis of Large Spermatocytes
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Cited by 6 •
2020
We present here an improved protocol for the whole-mount preparation and immunostaining of Drosophila testes, suitable for confocal microscopy and also allowing reproducible and reliable labeling. We illustrate this protocol by 3D representation and quantification of colocalization experiments on the large S4-S5 spermatocytes.
FAQs
PNGase F hydrolyzes the bond between the innermost N-acetylglucosamine and the asparagine side chain. This cleavage releases the attached oligosaccharide and converts the modified asparagine residue into aspartic acid. The resulting amino acid change provides a biochemical marker that the glycan-bearing site has undergone enzymatic deglycosylation.
Broad substrate specificity allows PNGase F to act on a wide range of glycoproteins rather than being limited to one particular protein substrate. This makes the enzyme useful for comparative glycosylation studies, in which researchers examine the same protein before and after carbohydrate removal to determine how N-linked glycans influence its measured properties or behavior.
Removing N-linked carbohydrates can produce detectable differences in a glycoprotein's molecular mass and electrophoretic mobility. Researchers can compare untreated and treated samples to assess whether carbohydrate removal changes the protein's apparent position during analysis. These comparisons also help reveal glycan-dependent effects on protein behavior without changing the protein sample being studied.
A typical workflow compares a glycoprotein before and after treatment with PNGase F. Researchers subject the samples to an analytical method, such as electrophoresis, immunoblotting, or mass spectrometry, and evaluate differences associated with glycan removal. The paired comparison connects observed changes in mass, mobility, or protein behavior with the presence of N-linked carbohydrates.
In electrophoresis, PNGase F treatment supports comparison of a glycoprotein's mobility before and after N-linked glycans are removed. Immunoblotting can then be used to examine the corresponding protein signal in the treated and untreated samples. Together, these approaches help determine whether carbohydrate removal produces a measurable shift while tracking the same protein.
Mass spectrometry can compare glycoprotein samples with and without their N-linked glycans, allowing researchers to assess glycan-dependent changes in molecular mass. PNGase F therefore supports structural characterization by separating observations associated with the protein from those contributed by attached carbohydrates. This use is relevant to biological studies of protein glycosylation and structure.