Cy5-Fuc Labeled sulfo-G2 (Cy5-sulfo G2)

Name: Cy5-Fuc Labeled sulfo-G2 (Cy5-sulfo G2)
Brand: R&D Systems
SKU: GL305

R&D Systems, part of Bio-Techne | Catalog # GL305

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Product Datasheet / COA / SDS

Key Product Details

Key Benefits:

Learn more about Fluorescent Glycan Labeling and Detection

Summary for Cy5-Fuc Labeled sulfo-G2 (Cy5-sulfo G2)

Formula	Fuc1Gal2Man3GlcNAc4Cy5S2
Predicted Mass	2775.35 Da
Formulation	Supplied in 20 mM Tris, pH 8.0
Stability & Storage	• 6 months from date of receipt, -20 to -70 °C as supplied. • 3 months, -20 to -70 °C under sterile conditions after opening. • Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

Applications

Used as a ligand for lectins and to study glycan protein interaction.
Used as a substrate for various glycosidase and glycosyltransferases.
Used as a substrate to study sulfatases.

Key Features:

Excitation at 649 nm and emission at 671 nm.
The fluorescent dye Cy5 is conjugated to the C6 position of the core-6 fucose.
Can be separated on 15-17% SDS-PAGE and directly visualized as a single band through the red channel of a fluorescent imager.
Linear response range for Cy5-labeled glycans can be from 10 fmol to 100 pmol, depending on the sensitivity of detection.

Related Reagents

Labeled Glycans

Click Chemistry

Enzymes and detection reagents

Sulfatases belong to a highly conserved family of enzymes that catalyze the hydrolysis of O- and N-sulfate esters from a variety of substrates. Among 18 human sulfatase genes identified, about 1/3 are found in lysosomes and serve to remove sulfate from glycosaminoglycans (GAGs), glycopeptides and glycolipids (GLs). The remaining sulfatases are found in microsomes, endoplasmic reticulum (ER), Golgi, at the cell surface, or secreted and are active against hydroxysteroids, heparin or unidentified substrates. Genetic deficiencies of individual sulfatases reveal the critical importance of these enzymes in the metabolism of specific forms of sulfate. Deficiencies of lysosomal sulfatases that act on GAGs and GLs account for mucopolysaccharidoses (MPS) and metachromatic leukodystrophy, respectively. Deficiencies in non-lysosomal sulfatases, arylsulfases C and E, cause X-linked ichthyosis (XLI) and chondrodysplasia punctata (CDPX), respectively.

Product Specifications for Cy5-Fuc Labeled sulfo-G2 (Cy5-sulfo G2)

Applications	Bioactivity
Source	Synthetic

Preparation & Storage

Shipping Conditions	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage	Protect from light. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

Background: Sulfatase-2/SULF2

Item	Value
Entrez Gene IDs	55959 (Human); 72043 (Mouse); 311642 (Rat)
Alternate Names	DKFZp313E091, EC 3.1.6, EC 3.1.6.-, EC 3.1.6.14, FLJ44488, FLJ90554, KIAA1247HSULF-2, MGC126411, SULF-2, SULF2, Sulfatase-2, Sulfatase2, extracellular sulfatase Sulf-2, hSulf-2, sulfatase 2

Assay Procedure

Sample Assay Protocol for testing activity of Sulfatases using Cy5-Fuc Labeled sulfo-G2/Cy5-sulfo G2 as a substrate.

Suggested input of Cy5-sulfo G2 in an assay separated on SDS-PAGE is from 0.01-1 pmol.

Protocols are guidelines. Parameters need to be optimized by end users, enzyme input and reaction time may need to be adjusted to accommodate specific activity of the enzyme.

Refer to Wu, ZL. et al. (2020) Glycobiology, 30:970.
https://academic.oup.com/glycob/article/30/12/970/5815178

Materials

Assay Buffer: 25 mM Tris, pH 7.5 (dependent on requirements of Sulfatase)
Sulfatase
15% SDS-PAGE
6X Gel Loading Dye
Fluorescent imager

Assay

Dilute Sulfatase 10 to 100 ng/µL in the Assay Buffer.
Dilute Cy5-sulfo G2 to 0.02 µM in Assay Buffer.
Mix 10 µL dilute Sulfatase and 10 µL of Cy5-sulfo G2 in a centrifuge tube.
Prepare a negative control by mixing 10 µL of Cy5-sulfo G2 with 10 µL of Assay Buffer.
Incubate the reaction and control at 37 °C for 1 hour.
Stop the reactions and controls by adding 4 µL of 6X Gel Loading Dye to each tube.
Load 12 µL of each of the above reactions and controls per well on a 15% SDS-PAGE and run down 80% length of the gel.
Image the gel using a fluorescent imager using the red channel for 10 seconds.

Final Assay Conditions Per Reaction

Sulfatase: 1 to 10 µg
Cy5-sulfo G2: 0.2 pmol

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