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

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (1)Q&A (14)References (6)

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MPO activity - ab111749
  • Functional Studies - Myeloperoxidase (MPO) Fluorometric Activity Assay Kit (ab111749)

Key features and details

  • Assay type: Enzyme activity
  • Detection method: Fluorescent
  • Platform: Microplate reader
  • Assay time: 30 min
  • Sample type: Cell culture supernatant, Tissue Extracts
  • Sensitivity: 0.0005 mU/well

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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

    > 0.0005 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

  • Notes

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests
    Assay Buffer WM 1 x 25ml
    Fluorescein Standard (1 mM) Yellow cap 1 x 50µl
    MPO Positive Control Purple cap 1 vial
    MPO Probe Red cap 1 x 200µl
    MPO Substrate Stock Blue cap 1 x 50µl
  • Research areas

    • Immunology
    • Innate Immunity
    • Macrophage / Inflamm.
    • Cardiovascular
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    • Fatty acid oxidation
    • Kits/ Lysates/ Other
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  • Function

    Part of the host defense system of polymorphonuclear leukocytes. It is responsible for microbicidal activity against a wide range of organisms. In the stimulated PMN, MPO catalyzes the production of hypohalous acids, primarily hypochlorous acid in physiologic situations, and other toxic intermediates that greatly enhance PMN microbicidal activity.
  • Involvement in disease

    Defects in MPO are the cause of myeloperoxidase deficiency (MPD) [MIM:254600]. MPD is an autosomal recessive defect that results in disseminated candidiasis.
  • Sequence similarities

    Belongs to the peroxidase family. XPO subfamily.
  • Cellular localization

    Lysosome.
  • Target information above from: UniProt accession P05164 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • 84 kDa myeloperoxidase
    • 89 kDa myeloperoxidase
    • EC 1.11.1.7
    • EC1.11.2.2
    • fj80f04
    • MPO
    • mpx
    • myeloid-specific peroxidase
    • Myeloperoxidase
    • Myeloperoxidase heavy chain
    • Myeloperoxidase light chain
    • PERM_HUMAN
    • wu:fj80f04
    see all

Associated products

  • Related Buffer

    • 10x Red Blood Cell (RBC) Lysis Buffer (ab204733)
  • Related Products

    • Anti-Myeloperoxidase antibody (ab45977)

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

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (6)

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

    ab111749 has been referenced in 6 publications.

    • Guenthart BA  et al. Regeneration of severely damaged lungs using an interventional cross-circulation platform. Nat Commun 10:1985 (2019). 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). PubMed: 29486477
    • You P  et al. Local level of TGF-ß1 determines the effectiveness of dexamethasone through regulating the balance of Treg/Th17 cells in TNBS-induced mouse colitis. Exp Ther Med 15:3639-3649 (2018). PubMed: 29545894
    • Kuwabara WMT  et al. Obesity and Type 2 Diabetes mellitus induce lipopolysaccharide tolerance in rat neutrophils. Sci Rep 8:17534 (2018). PubMed: 30510205
    • Roberts JS  et al. Opportunistic Pathogen Porphyromonas gingivalis Modulates Danger Signal ATP-Mediated Antibacterial NOX2 Pathways in Primary Epithelial Cells. Front Cell Infect Microbiol 7:291 (2017). PubMed: 28725637
    • Tang Z  et al. The role of bradykinin in lung ischemia-reperfusion injury in a rat lung transplantation model. Acta Cir Bras 31:807-812 (2016). PubMed: 28076504

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 15 Abreviews or Q&A

    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?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 01 2012

    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.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Oct 01 2012

    MPO activity of ovine lung tissue

    Good Good 4/5 (Ease of Use)
    Abreviews
    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)?

    Read More

    Abcam community

    Verified customer

    Asked on Jun 16 2014

    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.

    Read More

    Kevin Hanson

    Abcam Scientific Support

    Answered on Jun 16 2014

    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?


    Read More

    Abcam community

    Verified customer

    Asked on May 22 2014

    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.

    Read More

    Sam Washer

    Abcam Scientific Support

    Answered on May 22 2014

    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,

    Read More

    Abcam community

    Verified customer

    Asked on Feb 18 2013

    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.

    Read More

    Abcam Scientific Support

    Answered on Feb 18 2013

    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?

    Read More

    Abcam community

    Verified customer

    Asked on Feb 15 2013

    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.

    Read More

    Abcam Scientific Support

    Answered on Feb 15 2013

    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?

    Read More

    Abcam community

    Verified customer

    Asked on Feb 08 2013

    Answer

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

    I have updated our datasheet accordingly.

    Read More

    Abcam Scientific Support

    Answered on Feb 08 2013

    Question

    How will EDTA inhibit this assay?

    Read More

    Abcam community

    Verified customer

    Asked on Dec 10 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Dec 10 2012

    Question

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

    Read More

    Abcam community

    Verified customer

    Asked on Nov 29 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Nov 29 2012

    Question

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

    Read More

    Abcam community

    Verified customer

    Asked on Nov 20 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Nov 20 2012

    1-10 of 15 Abreviews or Q&A

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