Overview

  • Product name

    Monoamine oxidase B Activity Assay Kit ((MAOB Assay)
  • Detection method

    Colorimetric
  • Sample type

    Cell culture extracts
  • Assay type

    Enzyme activity
  • Species reactivity

    Reacts with: Human
  • Product overview

    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.

  • Notes

    Store Fluorophore and benzylamine at -80°C. Store all other components store at 4°C.

  • Platform

    Microplate reader

Properties

Images

  • 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 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.

    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.

Protocols

References

This product has been referenced in:

  • 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). Read more (PubMed: 30195017) »
See 1 Publication for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Answer

Thank you for your response.

It is important to emphasize that as the datasheet suggests this kit has been designed and tested on cell culture extracts but not specifically on serum samples. As long as MAOB (in serum) is detectable, you could make comparison between control samples and treated samples at different time points and/or at different concentrations.

If you need any further assistance, please do not hesitate to contact me.

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Answer

Thank you for contacting us.
This kit is intended for use in measuring MAOB in cell culture samples. This assay is an immunoassay, a recombinant or purified standard would not be necessarily comparable to a native sample for activity so we don't offer it. For example an MAOB recombinant protein is available from Abcam ab82944 however it is not classified for functional studies funcS therefore we are not sure if it will work. The best control is MAOB isolated from untreated cells lysate.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question
Answer

Thank you for your enquiry.

The antibodies used in this kit were specially made for the kit and have never been released for sale as a standalone product. Therefore, there was no need to create a clone number for them and the information you are looking for is not available.

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Answer

Thank you for your enquiry and your interest in our products.

The Monoamine oxidase B (MAOB) Specific Activity Assay Kit (ab109912) has been designed for cell culture extracts.

We would recommend using this kit in ‘profiling’ assay which means it is to be used when comparing the levels of protein in two samples – normal and unknown/treated.

We currently don’t have access to an acceptable AMOB protein that is sufficiently native and works in the kit. As soon as we have further information, we will update the on-line product datasheet accordingly.

For your interest there is also a MAOB activity/quantity assay ab109912 in our catalogue that may be of interest.

I hope this helps and if I can assist further, please do not hesitate to contact me.

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Answer

Thank you for your inquiry.

1. I can confirm that this assay does not deliver the result asa quantitative amount of MAOB. This kit can be used to quantitatively measure the different between two samples.

For example. Sample 1 is untreated and samples 2 is treated. the difference in MAOB can be measured.
If you would like to create your own standard curve, I suggest to MAOB in defined amounts.

2. Unfortunately, this kit is not tested and guaranteed for serum samples. I suggest to prepare a serial dilution of samples with assay buffer. This has to be optimised and is not guaranteed.

For more information please see the protocol booklet.

I am sorry I did not have positive answer to your questions on this occasion and wish you good luck with your research.

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Answer

Thank you for contacting Abcam. Based on the sequence homology between human and dog (97%), there is a high probability that kit would work in samples from dog but we cannot guarantee that it. If there is anything else I can help you with, please let me kn

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Answer

Thank you for contacting us! This assay is an immunoassay, a recombinant or purified standard would not be necessarily comparable to a native sample for activity so we don't offer it. For example an MAOB recombinant protein is available from Abcam ab82944 however it is not classified for functional studies funcS therefore we are not sure if it will work. The best control is MAOB isolated from untreated cells lysate. HepG2 cells are available from ATCC http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/tabid/452/Default.aspx?ATCCNum=HB-8065&Template=cellBiology MAOB is present in many human cell types we also have data, for example, with SHSY5Y cells. lastly, the data shown were in vitro (in-well), several exposure of drugs(including pargyline) in a dilution series to the isolated enzymes from normal untreated cells. The cells could be stimulated first then the enzyme isolated this may work -however we have not done this. The issue that may be faced here is that the extraction and dilution of the cell lysate into the assay buffer would change the local concentration of the pargyline thereby altering its effects on activity. I hope this information will be helpful. Should you have any other question please do not hesitate to ask.

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Answer

Thank you for your reply. I forwarded your question to the lab at Mitosciences, and they are not aware of a source of the active MAOB enzyme so they have not tested the kit with a recombinant standard. They recommend generating a standard curve with normal sample, demonstrated in the protocol. The data is generally reported as activity per mg or ug of normal sample. I hope this information is helpful, but please let me know if you have any further questions.

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Answer

Thank you for your phone call today. The activity and quantity curves in the ab109912 protocol were generated using HepG2 cell lysate, so it's not really a typical standard. For the curve with the triangle icon, the HepG2 cells were treated with pargyline prior to lysis. These representative curves show that the kit is compatible with this lysate in the range of 16-1000 ug/mL. I hope this information is helpful but please let me know if you have any further questions and I will be happy to help you.

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Answer

Thank you for your inquiry. These antibodies were generated using the shotgun immunization approach so we do not know the exact epitope. There was no peptide sequence used to immunize so unfortunately I cannot BLAST an immunogen sequence with pig. However I can say that the activity assay for MAOB is bovine (cow) crossreactive as tested by the laboratory (not shown on datasheet). Furthermore human, cow and pig have close similarity (91% identical in primary sequence).  Therefore, this assay has a good chance of working with pig but we wouldn't be able to guarantee it since we haven't yet tested this kit in pig. I hope this information helps. Please contact us with any other questions.

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