TThe 300 synthetic glycans featured in the Glycan-300 array consists of four groups of targets including the 100 glycans from the Glycan-100 array and an additional 80 N-glycans, 50 glycolipid glycans, and 50 human milk oligosaccharide glycans. The included glycans have been frequently identified as structures showing important binding functions in the literature.
Glycan Array 300
Glycans
Kit Components
- Dialysis Vials
- Labeling Reagent
- Stop Solution
- Glycan Array Glass Slide Assembly
- Sample Diluent
- 20X Wash Buffer I
- 20X Wash Buffer II
- Cy3 equivalent dye-conjugated Streptavidin
- Slide Washer/Dryer
- Adhesive device sealer
- Labeling Buffer
- Floating Dialysis Rack
Required Materials
- Detection antibodies of interest (For sandwich-based method only)
- Pipettors, pipet tips and other common lab consumables
- Orbital shaker or oscillating rocker
- Aluminum foil
- Distilled or de-ionized water
- 1.5ml Polypropylene microcentrifuge tubes
- Gene microarray scanner or similar laser fluorescence scanner
- Small plastic or glass containers
- KCl, NaCl, KH2PO4 and Na2HPO4 (For label-based method only)
- Beaker, stir plate and stir bar
Protocol Outline
- Dry the glass slide
- Block array surfaces
- Incubate samples (samples need to be biotinylated for the label-based approach)
- For the the sandwich-based principle, incubate with a detection antibody cocktail
For the label-based principle, incubate the labeled-streptavidin. - Incubate with Cy3 Equivalent Dye-Streptavidin
- Disassemble the glass slide
- Scan with a gene microarray laser scanner
- Perform densitometry and analysis
Storage
Upon receipt, all components of the Glycan Array 100 kit should be stored at -20°C. After initial use, remaining reagents should be stored at 4°C to avoid repeated freeze-thaw cycles and may be stored for up to 3 months (Labeling Reagent, Item B, should be prepared fresh each time before use). Unused glass slides should be kept at -20°C and repeated freeze-thaw cycles should be avoided (slides may be stored for up to 6 months). The entire kit should be used within 6 months of purchase.
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