Lysyl Oxidase Activity Assay试剂盒(Fluorometric) (ab112139)

概述

  • 产品名称Lysyl Oxidase Activity Assay试剂盒(Fluorometric)
  • 检测方法Fluorescent
  • 样品类型
    Cell culture supernatant, Cell culture extracts, Tissue Extracts
  • 检测类型Semi-quantitative
  • 灵敏度
    40 ng/well
  • 种属反应性
    与反应: Mouse, Rat, Human, Pig
    预测可用于: Mammal
  • 产品概述

    Lysyl Oxidase Activity Assay Kit (Fluorometric) (ab112139) provides a simple method to measure lysyl oxidase (LOX) activity in cell and tissue extracts, as well as physiological solutions. The assay uses a proprietary LOX substrate that releases hydrogen peroxide upon transformation by the LOX present in the sample. Hydrogen peroxide is in turn detected using our proprietary red fluorescence substrate for HRP-coupled reactions. This leads to increase in fluorescence that can be easily detected at Ex/Em = 540/590 nm in a fluorescence microplate reader.


    Please be aware that this assay is semi-quantitative as it does not contain a LOX standard for calibration. When a known concentration of LOX is used, the assay can detect activity from as low as 40 ng of lysyl oxidase in solution. The assay is much more sensitive than the other assays currently available in the market and its unique detection method eliminates the interference that occurs in certain biological samples.


    Visit our FAQs page for tips and troubleshooting.

  • 说明

    Lysyl oxidase (protein-lysine-6-oxidase, LOX, EC 1.4.3.13) is an extracellular copper-dependent enzyme that catalyzes formation of aldehydes from lysine residues in collagen and elastin precursors. These aldehydes are highly reactive and undergo spontaneous chemical reactions with other lysyl oxidase-derived aldehyde residues or with unmodified lysine residues. The chemical reactions result in cross-linking collagen and elastin, which is essential for stabilization of collagen fibrils and for the integrity and elasticity of mature elastin.

    Lysyl oxidase inhibition can cause osteolathyrism, while its upregulation by tumor cells may promote metastasis of an existing tumor, which makes LOX and important oncological target.

    The activity of Lysyl oxidase in biological samples is traditionally assessed by tritium release end-point assays using radio isotope labeled collagen or elastin substrates.

  • 经测试应用适用于: Functional Studiesmore details
  • 平台Microplate

性能

    相关产品

    应用

    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.

    应用 Ab评论 说明
    Functional Studies Use at an assay dependent concentration.

    Lysyl Oxidase Activity Assay Kit (Fluorometric) 图像

    • Relative LOX activity levels in mouse tissue. Tissue samples were prepared following assay protocol. Protein concentration was determined and samples were diluted 2-30 fold. LOX activity levels were measured after 15 minutes incubation in a fluorometric plate reader at Ex/Em = 535/587 nm. LOX activity levels are relative to background noise (blank control).

    • LOX activity fluorescence (RFU) values vs quantity (µg). Recombinant human LOX2 was serially diluted 3.35-0.04 (1/3) and activity was measured following assay procedure.

    • LOX activity absorbance (OD) values vs quantity (µg). Recombinant human LOX2 was serially diluted 3.35-0.04 (1/3) and activity was measured following assay procedure. Colorimetric detection is approximately 10-times lower than fluorometric.

    • Typical lysyl oxidase (LOX) dose response curve. Known amounts of LOX were added to wells and reaction was run following assay protocol. Fluorescence was measured on a solid black 96-well plate using a Gemini fluiorescence microplate reader (Molecular Devices). Activity from as low as 40 ng/mL of LOX can be detected after 30 minutes incubation.

    实验方案

    Lysyl Oxidase Activity Assay Kit (Fluorometric) (ab112139)参考文献

    This product has been referenced in:
    • Chang HM  et al. Activin A-induced increase in LOX activity in human granulosa-lutein cells is mediated by CTGF. Reproduction 152:293-301 (2016). Functional Studies ; Human . Read more (PubMed: 27530347) »
    • Ji F  et al. Hypoxia inducible factor 1a-mediated LOX expression correlates with migration and invasion in epithelial ovarian cancer. Int J Oncol 42:1578-88 (2013). Functional Studies ; Human . Read more (PubMed: 23545606) »

    See all 2 Publications for this product

    Product Wall

    Theoretically, azide should be compatible with ab112139 Lysyl Oxidase Activity Assay Kit (Fluorometric), though I am sorry we have not directly tested this so cannot provide data to confirm this.

    Compounds and buffer components that may interf...

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    The laborotory uses 500 uM BAPN from TCI, the cat# is A9754, and the cas# is 2079-89-2. In general the laboratory would recommend between 0.1 - 1 mM as most publications use BAPN concentrations in this range. Please see some example references attached.

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