IKK beta K-LISA Kit
from
EMD Millipore
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Product
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IKK beta K-LISA Kit
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Company
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EMD Millipore
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Price
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More Information
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View Company Product Page
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Detection
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Request Info
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Form
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96-well plate
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Target
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IKK beta
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Catalog Number
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CBA044-1KIT
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Method
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Colorimetric
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Original Item Name
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K-LISA™ IKKbeta Inhibitor Screening Kit
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Quantity
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96 Tests
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Range
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5-10 ng
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Sample Type
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Purified enzyme
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Description
The K-LISA™ IKKβ-Inhibitor Screening Kit is designed for rapid in vitro screening of IKKβ inhibitors. The assay is an ELISA-based activity assay that utilizes a 50-amino acid GST-IκBα fusion polypeptide substrate that includes the Ser32 and Ser36 IKKβ phosphorylation sites. The GST-IκBα substrate and IKKβ are incubated in the presence of IKKβ inhibitors in the wells of a Glutathione-Coated 96-Well Plate, which allows for substrate phosphorylation and capture in a single step. The phosphorylated GST-IκBα substrate is detected using an Anti-Phospho IκBα (Ser32/Ser36) antibody, followed by the HRP-Conjugate and color development with TMB Substrate. ELISA Stop Solution is used to stop the color development and the absorbance is read at 450 nm preferably with a reference wavelength of 540-600 nm. The absorbance is directly related to the level of IKKβ activity.
References
Greten, F.R., et al. 2004. Cell 118, 285.Baldwin, A.S., 2001. J. Clin. Invest. 107, 241.Yamamoto, Y. and Gaynor, R.B., 2001. J. Clin. Invest. 107, 135.Karin, M., et al. 2000. Annu. Rev. Immunol. 18, 621.Zandi, E., et al. 1997. Cell 91, 243.
Specifications/Features
Kit Contents:
The K-LISA™ IKKβ-Inhibitor Screening Kit is designed for rapid in vitro screening of IKKβ inhibitors. The assay is an ELISA-based activity assay that utilizes a 50-amino acid GST-IκBα fusion polypeptide substrate that includes the Ser32 and Ser36 IKKβ phosphorylation sites. The GST-IκBα substrate and IKKβ are incubated in the presence of IKKβ inhibitors in the wells of a Glutathione-Coated 96-Well Plate, which allows for substrate phosphorylation and capture in a single step. The phosphorylated GST-IκBα substrate is detected using an Anti-Phospho IκBα (Ser32/Ser36) antibody, followed by the HRP-Conjugate and color development with TMB Substrate. ELISA Stop Solution is used to stop the color development and the absorbance is read at 450 nm preferably with a reference wavelength of 540-600 nm. The absorbance is directly related to the level of IKKβ activity.