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    products/assay-kits/monoamine-oxidase-b-activity-assay-kit-maob-assay-ab109912.pdf

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Monoamine oxidase B Activity Assay Kit ((MAOB Assay) (ab109912)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (10)References (1)

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- Monoamine oxidase B (MAOB) Specific Activity Assay Kit (ab109912)
  • - Monoamine oxidase B (MAOB) Specific Activity Assay Kit (ab109912)
  • Sandwich ELISA - Monoamine oxidase B (MAOB) Specific Activity Assay Kit (ab109912)

Key features and details

  • Assay type: Enzyme activity
  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Sample type: Cell culture extracts

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Overview

  • Product name

    Monoamine oxidase B Activity Assay Kit ((MAOB Assay)
    See all Monoamine Oxidase B kits
  • 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

  • Storage instructions

    Please 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
  • Research areas

    • Neuroscience
    • Neurotransmitter
    • Biogenic Amines
    • Dopamine
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Parkinson's disease
    • Other
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Signal Transduction
    • Metabolism
    • Drug metabolism
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Other Metabolism Assay
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Drug metabolism
  • Alternative names

    • Amine oxidase [flavin-containing] B
    • MAO-B
    • MAOB
    • Monoamine oxidase type B

Images

  • - Monoamine oxidase B (MAOB) Specific Activity Assay Kit (ab109912)
    - Monoamine oxidase B (MAOB) Specific Activity Assay Kit (ab109912)
    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.
  • - Monoamine oxidase B (MAOB) Specific Activity Assay Kit (ab109912)
    - Monoamine oxidase B (MAOB) Specific Activity Assay Kit (ab109912)
    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.
  • Sandwich ELISA - Monoamine oxidase B (MAOB) Specific Activity Assay Kit (ab109912)
    Sandwich ELISA - Monoamine oxidase B (MAOB) Specific Activity Assay Kit (ab109912)
    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

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (1)

Publishing research using ab109912? Please let us know so that we can cite the reference in this datasheet.

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

Customer reviews and Q&As

Show All Reviews Q&A
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1-10 of 10 Abreviews or Q&A

Question

Thanks for your reply.

We will try to use this kit to assay serum samples and hope we can get some positive results. But I am a little confused about the method.

Now we have some samples collected from different time points. We will consider the samples before dose as normal samples and the samples after dose as treated samples, respectively.

Can we compare the MAOB level of treated samples with that of the normal samples at the same time point by directly loading these samples on the microplate and reading the response. I think that will be a qualitative assay, not a quantity assay. How do you think about it?Thanks ,

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Abcam community

Verified customer

Asked on Apr 25 2012

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.

Read More

Abcam Scientific Support

Answered on Apr 25 2012

Question


Inquiry: Hi, We are considering to purchase your Monoamine Oxidase B Enzyme Specific Activity Microplate Assay Kit (ab109912) and were also wondering if you carry the Monoamine Oxidase B which can work with this kit? Thank you!

Read More

Abcam community

Verified customer

Asked on Jan 02 2013

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.

Read More

Abcam Scientific Support

Answered on Jan 02 2013

Question

Can you tell me the clone number of the monoclonal antibodies used this kit?

Read More

Abcam community

Verified customer

Asked on Oct 01 2012

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.

Read More

Abcam Scientific Support

Answered on Oct 01 2012

Question

Dear Sir:

We ordered a Monoamine Oxidase B Enzyme Specific Activity Microplate Assay Kit to determine the MAOB level in the serum samples. Now, we have some questions on the performance of the kit as following:

1. How to perform the quantity analysis without the reference in the kit?

2. For the serum samples, will the pre-treatment be needed?

Thanks,

Read More

Abcam community

Verified customer

Asked on Apr 24 2012

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.

Read More

Abcam Scientific Support

Answered on Apr 24 2012

Question

尊敬的老师:

我公司客户购买了Abcam公司货号为ab109912的Monoamine oxidase B (MAOB) Specific Activity Assay Kit,在实验过程中遇到以下问题,请帮忙解决!

We ordered a Monoamine Oxidase B Enzyme Specific Activity Microplate Assay Kit to determine the MAOB level in the serum samples.

Now, we have some questions on the performance of the kit as following:

1. How to perform the quantity analysis without the reference in the kit?

2. For the serum samples, will the pre-treatment be needed?



谢谢!

林清

成都东方锐进科技有限公司






公司地址:成都市高新区益州大道天益街38号理想中心4栋1007室
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FAX:028-85141807
mobile:15008246179
em:linqing@orstc.com



http://www.orstc.cn/

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Abcam community

Verified customer

Asked on Apr 23 2012

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.

Read More

Abcam Scientific Support

Answered on Apr 23 2012

Question

Hello,   Do you think the kit for measuring Monomamine oxidase B (MAOB) would also work in dog samples?    

Read More

Abcam community

Verified customer

Asked on Dec 05 2011

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

Read More

Abcam Scientific Support

Answered on Dec 05 2011

Question

I have read that you do not offer a material in the kit to use as a standard in the afore mentioned MAOB assay.   Could you please clarify  where you attained the Hepg2 cells you did use in the assay?  Were they from ATCC?  If so what catalogue #?   Additionally, I read that pargyline was use to stimulate the cells prior to using them as materials for the standard curve. Could you share that procedure?   Did you use the same materials for controls?   Do you have any historical data for intra or inter-assay precision?   Thank you and have a great day.  

Read More

Abcam community

Verified customer

Asked on Aug 22 2011

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.

Read More

Abcam Scientific Support

Answered on Aug 22 2011

Question

What would the manufacturer recommend that a customer use for a standard for the activity and the quantity of the enzyme? Do they (or you) know of a quantified recombinant MAO-B enzyme that could be used for both activity and quantity? Have they tried using a recombinant for this purpose?  

Read More

Abcam community

Verified customer

Asked on Jul 22 2011

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.

Read More

Abcam Scientific Support

Answered on Jul 22 2011

Question

What standard was used to generate activity and quantity curves, and is the standard available for sale?

Read More

Abcam community

Verified customer

Asked on Jul 20 2011

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.

Read More

Abcam Scientific Support

Answered on Jul 20 2011

Question

What was the immunogen used to raise this antibody in this kit? Do you expect this kit to cross-react with pig?

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Abcam community

Verified customer

Asked on Jul 19 2011

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.

Read More

Abcam Scientific Support

Answered on Jul 19 2011

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