IKKbeta Human Other Protein Kinase Enzymatic Radiometric LeadHunter Assay [10 μM ATP] - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Nuclear factor kappa B (NF-kB) plays a critical role in the inducible expression of a large number of genes involved in immune, inflammatory and apoptotic processes. NF-kB is associated with IkB in cytoplasm, which inhibits NF-kB activity. IkB kinase (IKK), which consists of at least three proteins (IKK-1/a, IKK-2/b, and IKK-3/g), phosphorylates IkB and mediates IkB degradation. Once IkB is removed, NF-kB is free to enter the nucleus of the cell and transcriptionally activate its targeted genes.
Family:
Kinase
Sub Family:
Other Protein Kinase
Class:
Protein Ser/Thr
Protein Name:
IKKbeta
Uniprot Number:
O14920
Protein Aliases:
IKK-B|IKK-beta|IkBKB|I-kappa-B kinase 2|IKK2|Nuclear factor NF-kappa-B inhibitor kinase beta|NFKBIKB
Gene Name:
IKBKB
Gene ID:
3551
Gene Aliases:
IKK2|NFKBIKB|IKK-beta|IKKB
Accession Number:
NM_001190720
Organism:
Human
Construct Details:
Full Length, Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Radiometry
Measured Response:
Scintillation
Testing Information
Procedure Summary:
Human recombinant protein kinase IKK-2 expressed in Sf21 insect cells is used. Test compound and/or vehicle is preincubated with 500 ng/ml enzyme‡ in modified MOPS buffer pH 7.2 for 15 minutes at 37⁰C. The reaction is initiated by addition of 20 μM IKKtide, 10 μM ATP and 0.25 μCi [γ32P]ATP for another 30 minute incubation period and terminated by further addition of 3% H3PO4. An aliquot is removed and counted to determine the amount of [32P]IKKtide formed. Compounds are screened at 10 μM.
Substrate:
20 µM IKKtide
ATP Concentration:
10 uM
Incubation:
30 min at 37°C
Control Inhibitor:
Staurosporine
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
10
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Oncology/Immuno-Oncology
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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