The Raybiotech lectin array uses standard glass slides each spotted with 8 wells of identical lectin arrays. Each lectin, together with the positive controls is arrayed in duplicate. The slides each come with a 8-well removable gasket which allows for the process of 8 samples using one slide. Four slide slides can be nested into a tray, which matches a standard microplate and allows for automated robotic high throughput process of 32 arrays simultaneously. The RayBiotech lectin array provides a powerful new tool for glycosylation determination, drug discovery and biomarker development; all with limited samples volumes required.
- High sensitivity and specificity
- Low sample volume (10-100 µl per array)
- Large dynamic range of detection
- Compatible with most sample types
- Test 8 samples on each slide
- Suitable for high-throughput assays
AIA, Aleuria aurantia, Allium sativum, Amaranthus caudatus, Anguilla anguilla, Bauhinia purpurea, Concanavalin A, Datura stramonium, Dolichos biflorus, Erythrina cristagalli, Eunonymus europaeus, Galanthus nivalis, Griffonia (Bandeiraea) simplicifolia I, Griffonia (Bandeiraea), Hippeastrum hybrid, Jacalin, Lens culinaris, Lotus tetragonolobus, Lycopersicon esculentum, Maackia amurensis I, Maclura pomifera, Narcissus pseudonarcissus, Peanut, Phaseolus lunatus, Phaseolus vulgaris, Erythroagglutinin, Leucoagglutinin, Pisum sativum, Psophocarpus, Sambucus nigra I, Sambucus nigra II, Solanum tuberosum, Sophora japonica, Soybean, Ulex europaeus I, Ulex europaeus II, Urtica dioica, Vicia faba, Vicia villosa, Wheat Germ, Wisteria floribunda
After biotinylation of HRP we can determine what kind of glycans are conjugated to HRP by using the RayBiotech lectin array. Lectins, GNA, HHA, NPA showed strong signals after incubation with 0.33 ug/mL Biotin-HRP followed detection by streptavidin-fluorescence-dye (Figure A, B and C). The fluorescence signals from GNA, HHA and NPA can be blocked in a concentration depend manner by HRP itself (Figure A and B), which means that these fluorescence signals were generated based on the binding between HRP and the three lectins. As we know, GNA, HHA and NPA lectins specifically bind to mannose which indicates that HRP contains mannose. By adding increasing amount of mannose, the signal from GNA, HHA and NPA can be reduced (Figure A and C). The reduction signals from increasing concentrations of mannose confirms that HRP protein contains mannose in its glycocalyx. Lectin VVA binds to the streptavidin-fluorescence dye.
Using the lectin array, we can discover the different glycoprotein profiles of the serum samples or cell lysates from patient cohorts versus a control group. Below images showed the profiles of the glycans from serum samples detected by Biotin-anti-human IgG and Fluorescence dye-streptavidin.
- Identify and profile the glycans in their samples
- Determine whether their biomarker of interest has glycan moieties
- Find specific glycan binding ligands in biological samples
Quantitative analysis of lectin-glycoprotein interactions. Example: a concentration series of glycoproteins detected with the lectin array could reveal concentration dependent effects of lectin-glycan binding.
Determine the profile of bacterial cell-surface glycans. Example: Cell lysate from bacteria can be Biotinylated and hybridized to the lectin array. Analysis of the binding pattern and correlation with the known carbohydrate-binding specificities of the lectins can determine the glycans on the cell membrane.
- Lectin Array Glass Slide/s
- Sample Diluent
- 20X Wash Buffer I
- 20X Wash Buffer II
- Cy3 equivalent dye-conjugated Streptavidin
- Slide Washer/Dryer
- Adhesive device sealer
Other Materials Required
- For sandwich-based approach, detection antibody cocktail
- 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
- 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
Upon receipt, all components of the Raybiotech Lectin Array 40 kit should be stored at -20°C. Once thawed, the glass slide and Cy3 equivalent dye-conjugated Streptavidin should be kept at –20°C and all other components may be stored at 4°C. The entire kit should be used within 6 months of purchase.