Proteasome Human Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Proteasome (EC 3.4.99.46) is the main enzyme of the ATP-dependent ubiquitin conjugate and ubiquitin-independent pathways of a non-lysosomal proteolytic system in all eukaryotic cells. The purpose is to control numerous physiological processes, including signal transduction, DNA repair, chromosome maintenance, transcriptional activation, cell cycle progression, cell survival and certain immune cell functions. This is in addition to its more established roles of removal of misfolded, damaged and effete proteins. The complete 26S proteasome consists of a 20S proteolytic core complex, which is crowned at one or both ends by 19S regulatory complex. Proteasome inhibitors may be useful in the treatment of cancer, vascular and pulmonary inflammation. Inhibitors may also result in the overproduction and secretion of triglyceride-rich lipoproteins, suppress the inhibition of the glucose transporter GLUT-4 activity contributing to insulin resistance and diabetes and also increase levels of oxidative damage in neurodegenerative diseases.
Family:
Ubiquitin Modifying
Sub Family:
Proteasome
Organism:
Human
Construct Details:
Endogenous
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrofluorimetry
Measured Response:
Fluorescence
Testing Information
Procedure Summary:
Human erythrocyte 20S proteasome is used. Test compound and/or vehicle is preincubated with 2.5 μg/ml enzyme‡ in modified HEPES buffer pH 7.5 for 15 minutes at 37°C. The reaction is initiated by addition of 10 μM Suc-Leu-Leu-Val-Tyr-AMC for another 30 minute incubation period. Determination of the amount of AMC formed is read spectrophotometrically. Compounds are screened at 10 μM.
Substrate:
10 µM Suc-LLVY-AMC
Incubation:
30 min at 37°C
Control Inhibitor:
Lactacystin
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:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
RELATED SOLUTIONS
AdverseReactionEnzymes SafetyScreen Panel - TW
Item: PP90
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