Products

CBGlyco™ IP Kit (Glyco-030CL)

IP Kit provides components for effective immunoprecipitation (IP and co-IP) based on small-scale, covalent, affinity-resin immobilization of antibodies and other glycoproteins via oxidized sugar groups.      

Specifications
Product Size
25 reactions
Technique
Immunoprecipitation (IP)
Assay Type
Affinity Purification
Target Specificity
Not Target-Specific
For Use With (Equipment)
Microcentrifuge
Contents & storage
Sufficient For: 25 IP/Co-IP reactions, each using 20 µL antibody-coupled resin
• UltraLink Hydrazide Resin, 0.5 mL
• Coupling Buffer, 50 mL
• Aniline, 0.2 mL
• Sodium meta-periodate, 0.5 g
• Wash Buffer, 60 mL
• Zeba Spin Desalting Columns,0.5 mL, 25 columns
• IP Lysis/Wash Buffer, 50 mL
• PBS Pack (makes 500 mL), 1 pack
• Elution Buffer, pH 2.8, 50 mL
• Lane Marker Sample Buffer, Non-reducing, (5X), 5 mL
• Spin Columns-Screw Caps, 25 columns
• Collection Tubes, 2 mL, 100 tubes

Store at 4°C.
BACKGROUND
Description
The Kit uses hydrazide-activated UltraLink Resin to immobilize and perform 25 IP or Co-IP assays with different antibodies or glycoprotein ligands. The method is ideal for polyclonal antibodies, which typically have abundant carbohydrates (glycosylation) on their Fc portions. Monoclonal antibodies that contain adequate carbohydrates are also effective. Antibodies immobilized by this hydrazide-to-carbohydrate method have unobstructed antigen-binding sites and optimal purification capability. The prepared glycoprotein or antibody columns can be regenerated and reused at least five times for immunoprecipitation, co-immunoprecipitation or pull-down affinity-purification without significant loss in binding capacity.
Kit contains resin, buffers and reagents for immobilization; desalting columns; IP lysis/wash and elution buffers for immunoprecipitation; microcentrifuge spin columns and collection tubes; and loading buffer for SDS-PAGE
The IP Kit method results in covalent attachment of oxidized sugar groups of an IP antibody (or any other glycoprotein with oxidizable carbohydrate groups) to the hydrazide resin. The antibody resin is then incubated with the sample that contains the protein antigen of interest, allowing the antibody-antigen complex to form. After washing to remove non-bound components of the sample, the antigen is recovered by dissociation from the antibody with elution buffer supplied in the kit. The entire procedure is performed in a microcentrifuge spin cup, allowing solutions to be fully separated from the resin with a brief centrifugation. Only the antigen is eluted by the procedure, enabling it to be identified and further analyzed without interference from antibody fragments.

For lab research use only, not for diagnostic, therapeutic or any in vivo human use.

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