Lysyl Oxidase Activity Assay Kit (Fluorometric) (ab112139)

Overview

  • Product nameLysyl Oxidase Activity Assay Kit (Fluorometric)
  • Sample type
    Cell culture extracts, Adherent cells, Suspension cells
  • Assay typeDirect
  • Sensitivity
    40 ng/well
  • Species reactivity
    Reacts with: Human, Pig
    Predicted to work with: all Mammals
  • Product overview

    ab112139 offers a sensitive fluorescent assay for detecting the activity of lysyl oxidase. It utilizes a proprietary LOX substrate that releases hydrogen peroxide detected using our proprietary red fluorescence substrate in HRP-coupled reactions. This method allows the detection of sub ng/mL lysyl oxidase and is much more sensitive than the currently available assays. It eliminates the interference that occurs in some biological samples and can be readily used to detect lysyl oxidase activity in cell extracts or solutions. Its signal can be easily read by either a fluorescence microplate reader at Ex/Em = 540/590 nm or an absorbance microplate reader at ~576 nm.

    Visit our FAQs page for tips and troubleshooting.

  • Notes

    ab112139 should be stored desiccated.

    Please NOTE that a protein standard is not provided with this kit.

  • Tested applicationsFunctional Studiesmore details

Properties

    Applications

    Our Abpromise guarantee covers the use of ab112139 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Abreviews Notes
    Functional Studies Use at an assay dependent concentration.

    Lysyl Oxidase Activity Assay Kit (Fluorometric) images

    • Lysyl Oxidase Activity in cell lysates, O.D. values measured colorimetrically.

    • Lysyl Oxidase Activity in cell supernatants, O.D. values measured colorimetrically.

    • Lysyl Oxidase Activity in mouse tissue lysates, RFU values measured fluorometrically.

    • Lysyl Oxidase Activity in mouse tissue lysates, O.D. values measured colorimetrically.

    • Lysyl Oxidase RFU values vs quantity (µg) measured fluorometrically.

    • Lysyl Oxidase O.D. values vs quantity (µg)  measured colorimetrically.

    • ab112139 used to measure Lysyl oxidase dose response on a solid black 96-well with ab112139. As low as 40 ng of lysyl oxidase can be detected with 30 minutes incubation (n=3). The insert shows the low levels of lysyl oxidase detection.

    Protocols

    References for Lysyl Oxidase Activity Assay Kit (Fluorometric) (ab112139)

    ab112139 has not yet been referenced specifically in any publications.

    Product Wall

    Please note that this kit is not an ELISA kit but an activity assay kit.

    With regards to the standard used, when the kit was first developed, the lab used isolated LOX from pig heart, which is not commercial available. The kit is now tested ...

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    Frozen samples are suitable for use with this kit.

    For cell lysates, the cells should be collected and then homogenized in PBS. After spinning down the cells, the supernatant will be used in the assay. 50 uL of supernatant will be added per well of the assay plate, and follow step C.1. in the protocol ...

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    I can confirm that Solid Black micro plate was used in experiment with top-reading instrument.

    The clear bottom black and solid black plates can be selected based on experimental set up and instrument.


    1. Should one use fresh or frozen samples? --Both are OK as long as there is LOX in the sample
    2. Are tissue extracts OK?--Yes.


    1) Regarding the standard serial dilutions, you are correct it should be 0.04 ug to 4 ug. This has been updated in our online protocols, but older copies of the protocol may still incorrectly say 0.04 ng. Please make the standards starting with 0....

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    I contacted the scientists in the lab about this question, and we have not tested the kit with samples containing protease inhibitors, but theoretically the kit should be compatible with protease inhibitors. It should be fine to add these to your ...

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    Thank you for contacting us.

    The LOX activity is expressed as Fluorescent RFU units. Please check the following publication for reference;

    http://www.molecular-cancer.com/content/9/1/206

    I hope this information is helpful to ...

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    We do not have precise data for the sensitivity of colorimetric measurement. However, the sensitivity will decrease at least 30-100 times if using colorimetric measurement in theory.

    This product was not in our catalogue, however I have contacted the lab and have got their approval for adding the HRP substrate to the catalogue. The ab number is ab155473 and costs €‘125.

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"