Monoamine oxidase B Activity Assay Kit ((MAOB Assay) (ab109912)
Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Cell culture extracts
Product nameMonoamine oxidase B Activity Assay Kit ((MAOB Assay)
See all Monoamine Oxidase B kits
Sample typeCell culture extracts
Assay typeEnzyme activity
Species reactivityReacts with: Human
ab109912 (MS747) is a novel assay that uses a high affinity monoclonal capture antibody to selectively isolate MAOB from all other peroxidases and oxidases (including MAOA) in a tissue or cultured cell sample. After isolation and subsequent measurement of the enzyme's functional activity, the quantity of isolated MAOB is measured in the same well by adding a second monoclonal detector antibody, which is quantified using a colorimetric label (HRP). Both reactions take place in time-dependent manners proportional to the amount of enzyme captured in each well. By combining activity and quantity measurements, the enzyme's relative specific activity can be determined. Specific activity is useful for measuring up or down regulation of activity by site-specific modification or damage, and in response to specific inhibitors.
Store Fluorophore and benzylamine at -80°C. Store all other components store at 4°C.
Storage instructionsPlease refer to protocols.
Components 1 x 96 tests 100X Benzylamine Substrate 1 x 0.25ml 100X Detector Antibody 1 x 0.125ml 100X HRP Label 1 x 0.125ml 10X Blocking Solution 1 x 10ml 12-channel reagent reservoirs 1 unit 20X Wash Buffer 1 x 25ml 500X Fluorophore 1 x 50µl 500X Peroxidase 1 x 50µl 96-well Microplate (12 strips) 1 unit Extraction Buffer (ab260490) 1 x 15ml HRP Development Solution 1 x 12ml
- Amine oxidase [flavin-containing] B
- Monoamine oxidase type B
Figure 2. MAOB is selectively inhibited by selegiline and pargyline, but not clorgyline. In this example, raw data was exported to Graph Pad Prism for 4-parameter fit analysis and IC50 determination.
Figure 1. With a HepG2 cell lysate MAOB activity was clearly measurable in the 16-1000 µg/ mL range and quantity in the range 1-1000 µg/mL. The MAOB specific inhibitor pargyline inhibited activity 90% while not affecting quantity.
Abcam's protein quantity microplate assays use the well-established sandwich ELISA format, whereby capture and detector antibodies are used to immobilize and then quantify a target protein or enzyme. All of our microplate assays utilize our highly-validated immunocapture antibodies, which are able to capture large, multi-subunit enzyme complexes in their fully intact state. Capture antibodies are pre-coated in the wells of premium Nunc MaxiSorp™ modular microplates, which can be broken into 8-well strips. After the target has been immobilized in the well, a second monoclonal antibody, against a different epitope on the target, is added to the well. This detector antibody is either directly labeled with biotin, or a biotin-labeled goat anti-mouse secondary is added. Substrate plus HRP or AP conjugated to streptavidin provide a colorimetric signal that is readable by any plate readers capable of standard ELISA absorbance measurements.
Datasheets and documents
ab109912 has been referenced in 1 publication.
- Hong GU et al. Inflammatory mediators resulting from transglutaminase 2 expressed in mast cells contribute to the development of Parkinson's disease in a mouse model. Toxicol Appl Pharmacol 358:10-22 (2018). PubMed: 30195017