Address: 45-1 Ramsey Road, Shirley, NY 11967, USA    USA: 1-631-559-9269  1-631-448-7888   Fax: 1-631-938-8127  Europe: 44-207-048-3343   Email: info@creative-biomart.com
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Insulin Glargine Bioassay Kit

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Cat.No.:
Kit-1604
Product Overview:
Bioassay kits provide a robust, highly reproducible and easy-to-use, cell-based functional assay to study receptor activity through a chemiluminescent output. The bioassay kits contain the reagents needed for a functional assay including ready-to-assay cells, plating, dilution & detection reagents, positive control agonist and assay plates. The ready-to-assay kits are designed to accelerate and simplify development of a QC potency assay or a NAb assay for biologic drugs.
Size:
1000 dp (10 x 96-well)/200 dp (2 x 96-well)
Description:
These assays utilize Enzyme Fragment Complementation (EFC) technology, where the b-galactosidase (b-gal) enzyme is split into two fragments, ProLink (PK) and Enzyme Acceptor (EA). Independently these fragments have no b-gal activity; however, when forced to complement through protein-protein interactions, they form an active b-gal enzyme. In the assay approach for tyrosine kinases, the ProLink tag is fused to the C-terminus of the receptor. The EA is fused to a phosphotyrosine SH2 domain containing protein that is able to bind the activated RTK. Ligand-induced activation of the receptor results in receptor phosphorylation. The SH2-EA fusion protein binds the phosphorylated receptor, forcing complementation of PK and EA to form an active b-gal enzyme.
Target Species:
Human
Storage:
U2OS Insulin Bioassay Cells: Cells are shipped on dry ice and should arrive in a frozen state. To ensure maximum cell viability, thaw the vials as soon as possible upon receipt. If continued storage of the frozen vials is necessary, store as follows: • Short term (2 weeks or less): Store vials at -80°C immediately upon arrival. (DO NOT store at –80°C for more than 2 weeks). • Long term (greater than 2 weeks): Vials should ONLY be stored in the vapor phase of liquid nitrogen (LN2 ). Safety Warning: A face shield, gloves and lab coat should be worn at all times when handling frozen vials. The manufacturer of the cryovial recommends storing the vials in the vapor phase above the liquid N2 . Upon thawing, if LN2 is present in the cryovial, it rapidly converts back to its gas phase which can result in the explosion of the vial upon its removal from the LN2 tank. Bioassay Detection Kit: Store at -20°C. Once thawed, the detection reagents can be kept at 4°C for up to 4 days. For longer storage (up to the expiration date listed in the kit certificate of analysis), the reagent should be aliquoted and stored at -20°C until needed. Avoid multiple freeze-thaw cycles. To make aliquots suitable for testing one assay plate each, 1 mL of Detection Reagent 1 per aliquot should be dispensed and frozen down. 4 mL of Detection Reagent 2 per aliquot should be dispensed and frozen down separately. Do not mix the two reagents during aliquoting. AssayComplete Cell Plating Reagent 5: Once thawed, the Cell Plating Reagent can be stored at 4°C for up to 4 weeks. For longer storage (up to the expiration date listed in the kit certificate of analysis), the reagent should be aliquoted and stored at -20°C until needed. Avoid multiple freeze-thaw cycles. To make aliquots suitable for testing one assay plate each, 30 mL of reagent per aliquot should be dispensed and frozen down.Protein Dilution Buffer: Once thawed, the Protein Dilution Buffer can be stored at 4°C for up to 4 weeks. For longer storage (up to the expiration date listed in the kit certificate of analysis), the reagent should be aliquoted and stored at -20°C until needed. Avoid multiple freeze-thaw cycles. To make aliquots suitable for testing one assay plate each, 10 mL of reagent per aliquot should be dispensed and frozen down. This amount may vary depending on your stock sample concentrations and should be adjusted accordingly. Recombinant Human Insulin Control Agonist: Store at -20°C until ready to use (up to the expiration date listed in the kit certificate of analysis). Centrifuge the vial prior to opening to maximize recovery. When ready to use, reconstitute to a concentration of 10 mg/mL by adding 1 mL of 0.01N hydrochloric acid. Reconstituted ligand is stable for 1 week at 2-8°C. For longer storage (up to the expiration date listed on the kit Certificate of Analysis), it is recommended to store in working aliquots at -20 to -80°C. 96-well Tissue Culture Treated Plates: Store at room temperature.
Kit Components:
U2OS Insulin Bioassay Cells 2 vials 10 vials Bioassay Detection Kit 200 dp 1,000 dp • Detection Reagent 1 2 mL 10 mL • Detection Reagent 2 8 mL 40 mLAssayComplete Cell Plating Reagent 5 1 x 100 mL 2 x 100 mLProtein Dilution Buffer 1 x 50 mL 2 x 50 mL Control Agonist (Insulin) 1 vial 1 vial 96-well Clear-Bottom TC Treated, Sterile Plates w/lid 2 plates 10 plates


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Optional requirements on this protein +Expand
C-fusion   N-fusion  Non-tagged
His   GST  Fc  Others
<1.0 eu/μg   <0.1 eu/μg  <0.01 eu/μg  Not required
Monomer Isolation   Dimer Isolation   Not required
>80% by SDS-PAGE   >90% by SDS-PAGE  >95% by SDS-PAGE  Others
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45-1 Ramsey Road, Shirley, NY 11967, USA
USA: 1-631-559-9269  1-631-448-7888
Europe: 44-207-048-3343
FAX: 1-631-938-8127
EMAIL: info@creative-biomart.com

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