Overview

  • Product name

    Anti-KDM1/LSD1 antibody [3D12] - ChIP Grade
    See all KDM1/LSD1 primary antibodies
  • Description

    Mouse monoclonal [3D12] to KDM1/LSD1 - ChIP Grade
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, WB, ELISA, IP, ChIPmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment corresponding to Human KDM1/ LSD1.

  • Positive control

    • HeLa, NIH3T3, mouse P19 and human LNCaP. ICC/IF: HAP1-KDM1A cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab90966 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
ICC/IF Use a concentration of 1 µg/ml.
Flow Cyt Use 1-2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB 1/1000. Predicted molecular weight: 93 kDa.
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration.

Target

  • Function

    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.
  • Tissue specificity

    Ubiquitously expressed.
  • Sequence similarities

    Belongs to the flavin monoamine oxidase family.
    Contains 1 SWIRM domain.
  • Domain

    The SWIRM domain may act as an anchor site for a histone tail.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • Amine oxidase (flavin containing) domain 2 antibody
    • Amine oxidase, flavin containing, 2 antibody
    • AOF2 antibody
    • BHC110 antibody
    • BRAF35 HDAC complex protein BHC110 antibody
    • BRAF35-HDAC complex protein BHC110 antibody
    • BRAF35/HDAC complex, 110-kD subunit antibody
    • CPRF antibody
    • EC1 antibody
    • FAD binding protein BRAF35 HDAC complex, 110 kDa subunit antibody
    • Flavin-containing amine oxidase domain-containing protein 2 antibody
    • KDM 1 antibody
    • KDM1 antibody
    • Kdm1a antibody
    • KDM1A_HUMAN antibody
    • KIAA0601 antibody
    • LSD 1 antibody
    • LSD1 antibody
    • Lysine (K) specific demethylase 1 antibody
    • Lysine (K) specific demethylase 1A antibody
    • Lysine demethylase 1A antibody
    • Lysine specific histone demethylase 1 antibody
    • Lysine specific histone demethylase 1A antibody
    • Lysine-specific demethylase 1 antibody
    • Lysine-specific demethylase 1A antibody
    • Lysine-specific histone demethylase 1A antibody
    see all

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: KDM1 / LSD1 knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: Jurkat cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab90966 observed at 110 kDa. Red - loading control, ab181602, observed at 37 kDa.


    ab90966 was shown to specifically recognize KDM1/LSD1 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when KDM1/LSD1 knockout samples were used. Wild-type and KDM1 / LSD1 knockout samples were subjected to SDS-PAGE. ab90966 and ab181602 (loading control to GAPDH) were diluted to 1/500 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.

  • ab90966 staining KDM1A in wild-type HAP1 cells (top panel) and KDM1A knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab90966 at 1μg/ml concentration and ab202272 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

  • Ab90966 was tested for immunostaining (upper panel) and IP-Western (lower panel). For immunostaining analysis, HeLa cells were transfected with a shLSD1 plasmid also encoding GFP for two days followed by immunostaining using ab90966. For IP-Western analysis, HeLa nuclear extracts (~200ug for each reaction) were immunoprecipitated with or without 0.5ug LSD antibody followed by Western analysis at a dilution of 1:50.
  • Overlay histogram showing HeLa cells stained with ab90966 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab90966, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:

  • Zhou A  et al. Nuclear GSK3ß promotes tumorigenesis by phosphorylating KDM1A and inducing its deubiquitylation by USP22. Nat Cell Biol 18:954-966 (2016). Read more (PubMed: 27501329) »
See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thanks for your reply.

I have requested the immunogen information used to produce ab90966. I will forward it to you as soon as I receive the details.

We typically only provide the discount testing program codes prior to purchase of the product to be tested. However I can make an exception in this case as you were not aware of this promotion. The discount codeassociated with this promotion will provide you one free primary antibody of your choice after you have submitted your Abreview. Please see below for your discount code information, including expiration date of the code:

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DISCOUNT CODE: 100%ABR-T71FL
Expiration date: September 30 2012

I am very pleased to hear you would like to accept our offer and test ab90966 in Toxoplasma. This code will give you: 1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview for Toxoplasma and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for your enquiry.

We do not have small sample sizes available. In lieu of this, we do have a testing program which you may be intersted in. I have described it below using ab90966 as an example. But the promotion could be applied to any of the antibodies you have mentioned here. Please let me know if you would like to take advantage of the promotion with 1 or more antibodies or if you have any questions.

======

Thank you very much for your interest in ab90966.

To our knowledge, ab90966 has not been tested in Toxoplasma. Therefore, I can offer a discount off a future purchase if you buy ab90966 now, test it in Toxoplasma and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free primary antibody. If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab90966 in Toxoplasma Iwill then send a discount code. This code must be issued before purchasing ab90966 so please wait for my reply before ordering.

2. Purchase ab90966 either by phone, fax, or online (www.abcam.com).

3. Test it in Toxoplasma.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab90966 turns out to be unsuitable forToxoplasma you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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