Overview

  • Product name

    KDM1/LSD1 Activity Quantification Assay Kit (Fluorometric)
    See all KDM1 / LSD1 kits
  • Detection method

    Fluorescent
  • Assay type

    Enzyme activity
  • Assay time

    3h 00m
  • Species reactivity

    Reacts with: Mouse, Human, Plants
    Predicted to work with: Mammals, Other species
  • Product overview

    KMD1/LSD1 (Lysine Specific Histone Demethylase 1) is a histone demethylase that only acts on mono- or di-methylated lysines. KDM1/LSD1 acts as a co-repressor by mediating demethylation of H3 (meK4), a specific tag for epigenetic transcriptional activation.


    KDM1/LSD1 Activity Quantification Assay Kit (Fluorometric) (ab113460) is designed for the easy and fast fluorometric measurement of KDM1/LSD1 activity or inhibition. The antibody-based, immunospecific method directly detects KDM1/LSD1-converted demethylated products, rather than by-products, in a 96 stripwell microplate format.


    This kit may cross react with LSD2.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 48 tests 96 tests
    10X Wash Buffer 1 x 14ml 1 x 28ml
    8-Well Assay Strips (with Frame) 6 units 12 units
    Adhesive Covering Film 1 unit 1 unit
    Capture Antibody, 1000 µg/mL 1 x 5µl 1 x 10µl
    Detection Antibody, 400 µg/mL 1 x 6µl 1 x 12µl
    Fluoro Developer 1 x 10µl 1 x 20µl
    Fluoro Diluter 1 x 4ml 1 x 8ml
    Fluoro Enhancer 1 x 10µl 1 x 20µl
    LSD1 Assay Buffer 1 x 4ml 1 x 8ml
    LSD1 Assay Standard, 50 µg/mL 1 x 10µl 1 x 20µl
    LSD1 Inhibitor Tranylcypromine, 1 mM 1 x 20µl 1 x 40µl
    LSD1 Substrate, 50 µg/ml 1 x 60µl 1 x 120µl
  • Research areas

  • Relevance

    Histone demethylase that demethylates both 'Lys-4' (H3K4me) and 'Lys-9' (H3K9me) of histone H3, thereby acting as a coactivator or a corepressor, depending on the context. Acts by oxidizing the substrate by FAD to generate the corresponding imine that is subsequently hydrolyzed. Acts as a corepressor by mediating demethylation of H3K4me, a specific tag for epigenetic transcriptional activation. Demethylates both mono- (H3K4me1) and di-methylated (H3K4me2)H3K4me. May play a role in the repression of neuronal genes. Alone, it is unable to demethylate H3K4me on nucleosomes and requires the presence of RCOR1/CoREST to achieve such activity. Also acts as a coactivator of androgen receptor(ANDR)-dependent transcription, by being recruited to ANDR target genes and mediating demethylation of H3K9me, a specific tag for epigenetic transcriptional repression. The presence of PRKCB in ANDR-containing complexes, which mediates phosphorylation of 'Thr-6' of histone H3 (H3T6ph), a specific tag that prevents demethylation H3K4me, prevents H3K4me demethylase activity of KDM1A. Demethylates di-methylated 'Lys-370' of p53/TP53 which prevents interaction of p53/TP53 with TP53BP1 and represses p53/TP53-mediated transcriptional activation. Demethylates and stabilizes the DNA methylase DNMT1. Required for gastrulation during embryogenesis. Component of a RCOR/GFI/KDM1A/HDAC complex that suppresses, via histone deacetylase (HDAC) recruitment, a number of genes implicated in multilineage blood cell development.
  • Alternative names

    • Amine oxidase (flavin containing) domain 2
    • AOF2
    • BHC110
    • BRAF35 HDAC complex protein BHC110
    • BRAF35-HDAC complex protein BHC110
    • FAD-binding protein BRAF35-HDAC complex, 110 kDa subunit
    • Flavin-containing amine oxidase domain-containing protein 2
    • KDM1
    • KIAA0601
    • LSD1
    • Lysine (K) specific demethylase 1
    • Lysine (K) specific demethylase 1A
    • Lysine specific histone demethylase 1
    • Lysine specific histone demethylase 1A
    • Lysine-specific histone demethylase 1
    • Lysine-specific histone demethylase 1A
    see all
  • Database links

Images

  • Demonstration of inhibitory effects of a KDM1/LSD1 inhibitor detected by ab113460. [KDM1/LSD1 concentration: 200 ng/well].
  • Demonstration of high sensitivity of KDM1/LSD1 activity assay achieved by using recombinant KDM1/LSD1 with ab113460.

Protocols

References

ab113460 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer

Thank you for contacting us.
The kit ab113472 is designed to detect DNA demethylase activity and will detect the this activity using a unique methylated DNA substrate. The detection is then colorimetric and can be measured by optical density measure at 450nm. I do not think that this is adapted for measuring LSD1 activity.
We do however have different other kits, which might correspond more to what you are looking for:
We do have the EpiSeeker KDM1/LSD1 activity quantification kit which comes either colorimetric (ab113459) or fluorometric (ab113460).  This kit does measure the activity of the LSD1 by using the di-methylated histone H3-K4 as a substrate for LSD1. The fluorimetric assay is read at Ex/Em = 530/590 nm.
https://www.abcam.com/index.html?datasheet=113459
https://www.abcam.com/index.html?datasheet=113460
Alternatively we do also have the EpiSeeker Histone Demethylase KDM1/LSD1 Activity Quantification Assay Kit ab113457. This kit does detect the histone demethylase by indirect detection. https://www.abcam.com/index.html?datasheet=113457
It might be interesting to note that we do have a website dedicated to the EpiSeeker products. Please feel free to browse this site as well. www.abcam.com/episeeker
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

Answer

Thank you for contacting Abcam. For ab113459 and ab113460, both kits directly detects KDM1/LSD1-converted demethylated products, rather than by-products, in a 96 stripwell microplate format. The main difference between the 2 are that ab113459, has a colorimetric detection systems, whereas, ab113460 has a fluorescent detections method. Depending on the equipment available in your lab, would determine which one of these you would choose. The ab113457 measures histone demethylase KDM1/LSD1 activity/inhibition from a broad range of species including mammalian cells/tissues, plants, and bacteria and this is an indirect detection method, compared to the above kits. For ab113456, this kit is ready-to-use and provides all the essential components needed for screening KDM1/LSD1 inhibitors, which are directly interacting with the enzyme or blocking the binding to its substrate. You can find more information about these products and how to choose the correct kit for your needs, by clicking on the following link: https://www.abcam.com/index.htmlpageconfig=resource&rid=14469&source=pagetrap&viapagetrap=episeeker#3 If there is anything else I can help you with, please let me know.

Read More

Question
Answer

Thank you for contacting Abcam. I have heard back from the lab about your inquiry as to whether you can use the products of your IP reaction with the kit ab113460. They have informed me the kit ab113460 can be used with IP product containing LSD1/antibody/beads complex. The assay can be performed according to the protocol of the kit with increased wash time ( additional 2-3 wash time) after incubation of the complex and substrate (Step 2f of the protocol), as the beads ( especially magnetic beads) could non-specifically bind to the wells that causes the antibody in the complex to bind to the detection antibody and generates a signal at the late steps. If there is anything else I can help you with, please do not hesitate to contact me.  

Read More

Answer

Thank you for contacting us. In answer to your question, the EpiSeeker KDM1/LSD1 Activity Quantification Assay Kit (ab113460) can be used to test LSD1 inhibitors both in vivo and in vitro. As you suggested, for in vivo testing, the cells can be incubated with the inhibitors (inhibitor compounds need to be capable of entering into the cells) for a defined time period. Nuclear extracts of these cells can then be used for the measurement of LSD1 activity with an untreated cell sample tested in parallel as a standard. The inhibitory effect can then be calculated by comparing the activity of the treated samples to that of untreated control. For in vitro testing, inhibitors can be directly incubated with the nuclear extract or purified LSD1 enzyme and then the level of inhibition calculated by comparing with untreated samples. 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

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up