Beta-Lactamase Bacteria Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
b-Lactamases, penicillin (cephalosporin) b-lactam amidohydrolases [EC3.5.2.6], are enzymes of bacterial origin which hydrolyze the C-N bond in the b-lactam ring of penicillin or cephalosporin. The inhibitors of b-lactamases, such as clavulanic or olivanic acids, are used clinically to enhance the actions of b-lactam antibiotics.
Family:
Cytosolic (Other)
Organism:
Bacteria
Construct Details:
Endogenous
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrophotometry
Measured Response:
Absorbance
Testing Information
Procedure Summary:
Bacterial b-Lactamase is used. Test compound and/or vehicle is preincubated with 0.132 μg/ml enzyme‡ in phosphate buffer pH 7.0 for 15 minutes at 25⁰C. The reaction is initiated by addition of 200 μM nitrocefin for another 60 minute incubation period. Determination of the amount of hydrolyzed nitrocefin formed is read spectrophotometrically. Compounds are screened at 10 μM.
Substrate:
200 μM Nitrocefin
Incubation:
60 min at 25°C
Control Inhibitor:
Tazobactam
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
DrugMatrixScreen LeadHunter Panel - TW
Item: PP89
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