Myeloperoxidase (MPO) Activity Assay Kit (Fluorometric) (ab111749)

Reviews (1)Q&A (14)References (5)

Overview

  • Product name

    Myeloperoxidase (MPO) Activity Assay Kit (Fluorometric)
    See all Myeloperoxidase kits

  • Detection method

    Fluorescent

  • Sample type

    Cell culture supernatant, Tissue Extracts

  • Assay type

    Enzyme activity

  • Sensitivity

    > 0.001 mU/well

  • Assay time

    0h 30m

  • Species reactivity

    Reacts with: Mammals, Other species

  • Product overview

    Myeloperoxidase (MPO) Activity Assay Kit ab111749 provides a rapid, simple and reliable fluorometric assay to study MPO activity.


    In the MPO assay protocol, MPO catalyzes the production of sodium hypochlorite (NaClO) from hydrogen peroxide (H2O2) and sodium chloride (NaCl). Subsequently, NaClO reacts stoichiometrically with the free radical sensor Aminophenyl fluorescein (APF) to generate fluorescein, whose fluorescence can be detected at Ex/Em = 485/525 nm.


    This MPO assay kit can be used to detect MPO activity as low as 0.5 µU per well.


    MPO assay protocol summary:
    - add fluoroscein standard to wells and analyze with a microplate reader
    - add samples to wells
    - add reaction mix
    - analyze every 2-3 mins with a microplate reader in kinetic mode for at least 30 mins

  • Platform

    Microplate reader

Properties

Images

  • MPO activity - ab111749
    MPO activity - ab111749

    a. Measurement of MPO activity in WBC lysate (0.1 μg) and MPO Positive Control (3 μL). b. MPO specific activity in WBC lysate.

  • Functional Studies - Myeloperoxidase (MPO) Fluorometric Activity Assay Kit (ab111749)
    Functional Studies - Myeloperoxidase (MPO) Fluorometric Activity Assay Kit (ab111749)
    Standard Curve using this kit protocol (Ex/Em = 485/525 nm)

Protocols

References

This product has been referenced in:

  • Guenthart BA  et al. Regeneration of severely damaged lungs using an interventional cross-circulation platform. Nat Commun 10:1985 (2019). Read more (PubMed: 31064987) »
  • Thomas AM  et al. Combined Inhibition of C5 and CD14 Attenuates Systemic Inflammation in a Piglet Model of Meconium Aspiration Syndrome. Neonatology 113:322-330 (2018). Read more (PubMed: 29486477) »
See all 5 Publications for this product

Publishing research using ab111749? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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Question



Inquiry: Dear Technical Support team: We are interested in measuring MPO activity in mouse liver tissue. I read that Myeloperoxidase (MPO) Activity Assay Kit (Fluorometric) (ab111749) is compatible with tissue extract. Is this kit compatible with snap frozen liver tissue or freshly harvested liver tissue? How does this kit distinguish MPO activity from that of other peroxidases in the tissue?

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Answer

Thank you for contacting us.

I would recommend using fresh tissue samples.

Other peroxidases will not be able to produce the same intermediates required for this assay. Therefore it is specific for MPO.


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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MPO activity of ovine lung tissue

 Good Good 4/5 (Ease of Use)
Abreviews
abreview image
Application: ELISA - MPO fluorometric assay
Samples: 10mg of homogenized lung tissue in 400uL of assay buffer
Loading amount: NEAT
Followed assay protocol to determine neutrophil infiltration in sheep lung tissue.
Standard curve: very high readings, out of range of neat samples - considered diluting standard to get within reference range
Positive control: was at the bottom end of the standard curve and used as a reference only to check the assay was working correctly

Ms. Gabriela Simonova

Verified customer

Submitted Sep 03 2014

Question

I have 20 different samples that I want to compare the activity for, so I need to normalize the amount of each sample I'm inputting. Would the assay buffer be compatible with a standard protein concentration measurement (eg. BCA assay)?

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Answer

The laboratory confirmed that the assay buffer will be compatible with a BCA assay, and this method of normalizing your samples' protein concentration would be appropriate in order to make activity level comparisons between your samples.

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Question

We are interested in the fluometric mpo activity assay (ab1117490). We were wondering whether it is it advisable to freeze the tissue from which the mpo activity will be analysed? If yes, do you freeze the tissue directly or do you first homogenize and then freeze the tissue?


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Answer


For peroxidase, fresh samples are the best. If needed the tissue can be quick-frozen in liquid nitrogen and stored at -80C. I would not advise making the lysate and then storing it for best results.

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Question

Hi,
I will be using your product ab111749 MPO activity assay kit for mouse organ samples.
Since we have many animals to sacrifice, we wonder if we can keep organ samples in freezer for using this MPO activity kit.
I am considering to necrospy and collect organ samples and homogenize samples with 4 volume of Assay Buffer as recommended in instructions,
then keep those samples in -80 degree till measure MPO activity.
Would you recommend to use those samples for measuring MPO activity at same day?
Can I keep samples in -80 as describe above?
Thank you,

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Answer

Thank you for your enquiry regarding ab111749.

We have always used with fresh samples only with this assay. We know of scientists who did store the homogenates in aliquots at -80°C for a short while before use but since we don’t ourselves have a comparative data measuring MPO in stored vs fresh samples, I cannot comment on if and how much of efficiency of detection is lost in freezing the samples.

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

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Question

Customer kindly contacted us regarding MPO activity assay kit (Fluorometric) with a couple questions.
1. Can tissue samples homogenized in assay buffer be frozen at -20 or -80? If so, what buffer conditions are appropriate (protease inhibitors, etc.)
2. In calculation step (Protocol step V. 5) V = volume of pretreated sample in the reaction well, which would be the sample volume in step V. 2 correct?

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Answer

The homogenized samples can be stored in aliquots at -80°C, w/o addition of any other components to the assay buffer. It would be preferable though to use fresh samples.
“V” is the volume of the sample you add to the wells in step 4.1.

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Question



‘Can this MPO assay kit (https://www.abcam.com/Myeloperoxidase-MPO-Activity-Assay-Kit-Fluorometric-ab111749.html) be used for rat tissue homogenates, or is it only for human samples?

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Answer

This kit can be used with all mammalian samples including rats.

I have updated our datasheet accordingly.

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Question

How will EDTA inhibit this assay?

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Answer

This is an enzymatic activity assay. Enzymes need some ions/cofactors to function and EDTA being a chelator will chelate these co-factors (specifically Mg++ and Ca++), leading to the enzyme becoming non-functional.

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Question

What is the concentration of the MPO positive control for my lot?

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Answer

Thank you for your inquiry and your patience with this reply.

I heard back from the supplying laboratory that the concentration of the positive control MPO does not vary from lot to lot. But unfortunately, they could not disclose the concentration for "proprietary reasons." I am sincerely sorry that we were not able to provide you with this information, but it seems as though the lab is unwilling and unable to release this information.

I understand that this is frustrating, but please let me know if we could help you in any other way.

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Question

Should a black plate with clear bottom wells or opaque bottom wells be used with this kit?

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Answer


For this assay kit, the client can either use a black opaque bottom plate or, for even more efficiency, use a white opaque bottom plate. One would not use a clear transparent plate.

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