• 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 glycosidases and glycosyltransferases.
Used as a scaffold for the synthesis of various glycan epitopes such as sialyl Lewis X structures.
Key Features and Benefits:
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 under a fluorescent imager.
Linear response range for Cy5-labeled glycans can be from 10 fmol to 100 pmol, depending on the sensitivity of detection.
Cy5-Fuc Labeled M1N1f as a substrate for recombinant MGAT2 assay. Cy5-Fuc labeled M1N1f (M1N1f) (Catalog # GL301) (0.2 pmol per reaction) was treated by a serial dilution of Recombinant Human N-Acetylglucosaminyltransferase2/MGAT2 Protein (Catalog # 10711-GT). The reactions were incubated 30 minutes at 37oC then heat inactivated, and further elongated with Recombinant Human B4GalT1 Protein (Catalog # 3609-GT) at 37oC for 20 minutes. Samples were separated on 15% SDS-PAGE and imaged with TCE staining (top panel) and fluorescent imaging (lower panel). Product 2 is visible with 6.25 ng of MGAT2, and its maximal level was achieved at 100 ng of the enzyme. The conversion of M1N1f to M2f by MGAT2 did not generate obvious mobility shift (data not shown). To overcome this issue, the substrate M1N1f and the product M2f were elongated by B4GalT1 to Product 1 and Product 2, respectively, to create an obvious gel mobility shift.
Product Specifications for Cy5-Fuc Labeled M1N1f
Source
Synthetic
Scientific Data Examples for Cy5-Fuc Labeled M1N1f
Cy5-Fuc Labeled M1N1f'
This substrate can be used as ligand for lectins and to study various glycosidases and glycosyltransferases. This can also be used as a scaffold for the synthesis of various glycan epitopes such as sialyl Lewis X structures. The fluorescent dye Cy5 is conjugated to the C6 position of the core-6 fucose.
Preparation & Storage
Storage
Store the unopened product at -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
Assay Procedure
Sample Assay Protocol for using Cy5-Fuc Labeled M1N1f/M1N1f as substrate for Recombinant Human N-acetylglucosaminyltransferase 2/MGAT2 assay
Protocols are guidelines. Parameters need to be optimized by end users. Suggested input of a Cy5-M1N1f in an assay separated on SDS-PAGE is from 0.01-1 pmol.
Materials
Assay Buffer: 25 mM Tris, 10 mM CaCl2, pH 7.5
Recombinant Human N-Acetylglucosaminyltransferase2/MGAT2 (rhMGAT2) (Catalog # 10711-GT)
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