Log In Sign up
The largest database of trusted experimental protocols

Glcnaz

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in United States

GlcNAz is a chemical compound used in biochemical research and analysis. It serves as a tool for studying glycosylation, a post-translational modification process in cells. GlcNAz can be incorporated into cellular glycans, enabling the visualization and tracking of glycosylated biomolecules. The product provides researchers with a versatile means to investigate the dynamics and functions of glycosylation in various biological systems.

Automatically generated - may contain errors

Lab products found in correlation

Galnaz, by Thermo Fisher Scientific (2 mentions) L azidohomoalanine aha, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) Ethylene glycol tetraacetic acid (egta), by Merck Group (1 mentions) Mannaz, by Thermo Fisher Scientific (1 mentions) Operetta high content imaging system, by PerkinElmer (1 mentions) Fv3000rs confocal microscope, by Olympus (1 mentions) Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (1 mentions) Prolong glass mounting reagent, by Thermo Fisher Scientific (1 mentions)

4 protocols using glcnaz

1

Visualizing Surface Proteins and Glycans

Check if the same lab product or an alternative is used in the 5 most similar protocols
For visualization of newly-synthesized proteins on the cell surface, neurons were incubated in methionine-free media subsequent to TNR epitope blocking: DMEM (4.5 mg/ml glucose, lacking pyruvate, methionine, glutamine, and cysteine; Life Technologies, Carlsbad, CA, USA) supplemented with 50 μM L-azidohomoalanine (AHA; Life Technologies), 812 μM MgCl2, 6.5 mM HEPES, 260 μM cysteine, 1:50 B27 (Gibco, Life Technologies), and 1:100 GlutaMAX (Gibco, Life Technologies). For visualization of surface glycans, 50 μM Click-IT™ GalNAz and/or GlcNAz (#C33365, C33367, Thermo Fisher Scientific, USA) were diluted directly into the neurons’ media subsequent to blocking, and remained throughout the experiment. Live, copper-free click labeling of surface AHA or glycans was performed after TNR labeling using strain-promoted azide-alkyne cycloaddition (SPAAC)68 . Neurons were incubated with 1:1000 dibenzocyclooctyne (DBCO) conjugated to AlexaFluor647 (#CLK-1302, Jena Bioscience, Germany) diluted in Ca2+/Mg2+-free Tyrode’s solution with 1 mM EGTA (#67-42-5; Merck, Germany).
Dankovich T.M., Kaushik R., Olsthoorn L.H., Petersen G.C., Giro P.E., Kluever V., Agüi-Gonzalez P., Grewe K., Bao G., Beuermann S., Hadi H.A., Doeren J., Klöppner S., Cooper B.H., Dityatev A, & Rizzoli S.O. (2021). Extracellular matrix remodeling through endocytosis and resurfacing of Tenascin-R. Nature Communications, 12, 7129.
+ Open protocol
+ Expand
2

Synthetic Carbohydrate Reagents for Biochemical Studies

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Peracetylated 2-Acetamido-1,3,6-tri-O-acetyl-4-deoxy-4-fluoro-D-glucopyranose (4F-GlcNAc) and 2-Acetamido-1,3,6-tri-O-acetyl-4-deoxy-4-fluoro-D-galactopyranose (4F-GalNAc) were from previous studies 20 (link), 27 (link). Peracetylated 1-(Naphthalen-2-ylmethanol) 2-Acetamido-3,4,6-tri-O-Acetyl-2-Deoxy-1-Thio-β-D-Glucopyranoside, GlcNAc-β-S-NAP (abbreviated SNAP), and corresponding peracetylated O-glycoside GlcNAc-β-O-NAP (abbreviated ONAP) were recently described 21 (link). Azido-derivatized carbohydrates ManNAz (N-azido acetyl-mannosamine tetraacylated), GalNAz (N-azido acetyl-galactosamine tetraacylated) and GlcNAz (N-azido acetyl-glucosamine tetraacylated) were purchased from ThermoFisher. Nucleotide-sugars and nucleotides were obtained from either Carbosynth (Compton, UK) or Sigma-Aldrich (St. Louis, MO). All other chemicals were from either ThermoFisher or Sigma unless otherwise mentioned.
del Solar V., Gupta R., Zhou Y., Pawlowski G., Matta K.L, & Neelamegham S. (2020). Robustness in glycosylation systems: Effect of modified monosaccharides, acceptor decoys and azido sugars on cellular nucleotide-sugar levels and pattern of N-linked glycosylation. Molecular omics, 16(4), 377-386.
+ Open protocol
+ Expand
3

Metabolic Labeling and Imaging of Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Sugar ManNAz, GalNAz, GlcNAz and Alexa Fluor 488 DIBO Alkyne (DIBO488) were purchased from ThermoFisher. Live cells were treated with 50 μM sugars at either 2, 6, 24 and 48 hours. Afterwards, cells were washed and were labelled with 15 μM DIBO488 and 5 μM COA-1 for 1 h at 37 °C, followed by counterstaining with nuclear tracker, DRAQ5. Cells were imaged using the Operetta High-Content Imaging System (PerkinElmer) with a 40× objective lens.
Alamudi S.H., Su D., Lee K.J., Lee J.Y., Belmonte-Vázquez J.L., Park H.S., Peña-Cabrera E, & Chang Y.T. (2018). A palette of background-free tame fluorescent probes for intracellular multi-color labelling in live cells †Electronic supplementary information (ESI) available: Experimental details, synthetic procedures, characterization data of compounds and supporting imaging. See DOI: 10.1039/c7sc04716a. Chemical Science, 9(8), 2376-2383.
+ Open protocol
+ Expand
4

Glycocalyx Quantification via Click-Chemistry

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were seeded on glass coverslips and treated with 40 µM of GlcNAz (Thermo Fisher Cat# PI88905) for 48 h. After three washes with cold DPBS (Gibco Cat# A1285801), cells were Cu-click conjugated with AlexaFluor-488-alkyne (Click Chemistry Tools Cat# 12771) and fixed as previously reported (83 ). Cells were mounted onto glass slides with ProLong glass mounting reagent (Thermo Fisher Cat# P36980). Images were acquired at 60× magnification on the Olympus FV3000RS confocal microscope.
The glycocalyx quantification was determined by defining a region of interest around individual cells and then measuring the intensity of the staining in each region using ImageJ software. Intensity plot of the cell membrane was generated and the width at half maximum intensity was considered as glycocalyx thickness.
Al-Nakouzi N., Wang C.K., Oo H.Z., Nelepcu I., Lallous N., Spliid C.B., Khazamipour N., Lo J., Truong S., Collins C., Hui D., Esfandnia S., Adomat H., Clausen T.M., Gustavsson T., Choudhary S., Dagil R., Corey E., Wang Y., Chauchereau A., Fazli L., Esko J.D., Salanti A., Nelson P.S., Gleave M.E, & Daugaard M. (2022). Reformation of the chondroitin sulfate glycocalyx enables progression of AR-independent prostate cancer. Nature Communications, 13, 4760.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!