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Lectin Array 95

Lectin Array 95

ArtNr	GA-Lectin-95-12
Hersteller	Raybiotech
Menge	12 Sample Kit
Quantity options	12 Sample Kit 24 Sample Kit 48 Sample Kit
Kategorie	Arrays Protein Arrays
Typ	Glycobiology Arrays
Specific against	other
Host	Hamster - Armenian
ECLASS 10.1	32161000
ECLASS 11.0	32161000
UNSPSC	41116126
Versandbedingung	Gekühlt
Lieferbar
Manufacturer - Applications	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
Number of Targets Detected	95
Compatible Sample Types	Cell Culture Supernatants Cell Lysates Plasma Serum Tissue Lysates
Design Principle	Label-basedSandwich-based
Method of Detection	Fluorescence Laser Scanner
Quantitative/Semi-Quantitative	Semi-Quantitative
Solid Support	Glass Slide
Kit Components	Dialysis Vials Labeling Reagent Labeling Buffer Stop Solution Lectin Array Glass Slide Assembly Sample Diluent20 X Wash Buffer I20 X Wash Buffer I I Cy3 equivalent dye-conjugated Streptavidin Slide Washer/ Dryer Adhesive device sealer Floating Dialysis Rack Manual
Other Materials Required	Detection antibodies of interest ( For sandwich-based method only) Orbital shaker Laser scanner for fluorescence detection Aluminum foil1.5ml Polypropylene microcentrifuge tubes K Cl, Na Cl, K H2 P O4 and Na2 H P O4 ( For label-based method only) Plastic or glass containers, beaker, stir plate and stir bar Pipettors, pipette tips, dd H2 O and other common lab consumables
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/Stability	Upon receipt, all components of the Raybiotech Lectin Array 95Â kit should be stored at -20°C. Once thawed, the glass slide and Cy3 equivalent dye-conjugated Streptavidin should be kept at Â–20°C and all other components may be stored at 4°C. The entire kit should be used within 6 months of purchase.
Additional Application Information	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.
Product Feature	High sensitivity and specificity Low sample volume (10-100 Âµl per array) Large dynamic range of detection Compatible with most sample types Test 12 samples on each slide Suitable for high-throughput assays
Application 1 - Detection of Glycans on a Purified Protein	See Image/s In this application, the RayBio Lectin Array 95 was used to detect specific glycosylations of purified Horseradish Peroxidase (HRP). Lectins BANLEC, BC2L-A, CALSEPA, GNA, HHA, NPA, PA-IIL, and PALa showed strong signals after incubation with 3.3 ug/mL Biotin-HRP followed by detection with streptavidin-fluorescence-dye (Figures A, B and C). The fluorescence signals from BANLEC, BC2L-A, CALSEPA, GNA, HHA, NPA, PA-IIL, and PALa were blocked in a concentration-dependent manner by HRP itself (Figures A and C), indicating that the signals were generated by lectin-HRP binding. These eight lectins are known to exhibit specific binding to mannose, which indicates that HRP contains mannose. After adding increasing amounts of mannose, the signals from BANLEC, BC2L-A, CALSEPA, GNA, HHA, NPA, PA-IIL, and PALa were reduced (Figures A and B).The reduction in signals from increasing concentrations of mannose confirms that HRP protein contains mannose in its glycocalyx. Additionally, the two lectins AAL and RS-FUC (fucose binding specificity) also showed strong interaction with HRP, which indicates the fucosylation of HRP. Overall, the results of the Lectin Array 95 were consistent with published literature regarding HRP glycosylation.
Application 2 - Profiling of a Serum Sample	See Image/s Using the lectin 95 array, we can discover the different glycoprotein profiles of serum samples, cell lysates, or purified glycoproteins. The images above show the profiles of the glycans from different types of samples including human serum, recombinant glycoproteins human HE4 and AFP, mouse TFF2, purified human IgG, and bacterial DE3 cell lysates detected by Biotin labeling and fluorescent dye-streptavidin.

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