Overview

  • Product name
    Anti-LSD2 / AOF1 antibody [EPR18508]
    See all LSD2 / AOF1 primary antibodies
  • Description
    Rabbit monoclonal [EPR18508] to LSD2 / AOF1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human LSD2/ AOF1 aa 50-150. The exact sequence is proprietary.
    Database link: Q8NB78

  • Positive control
    • WB: NIH/3T3, C6, RAW 264.7 and PC-12 cell lysates; Human fetal brain and fetal heart lysates; Mouse thymus lysate. ICC/IF: HeLa and A431 cells. IP: K562 whole cell lysate.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab193080 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
Flow Cyt Use at an assay dependent concentration.

Purified format.

ICC/IF 1/500.
IP 1/80.
WB 1/2000. Detects a band of approximately 92 kDa (predicted molecular weight: 92 kDa).

Target

  • Function
    Histone demethylase that demethylates 'Lys-4' of histone H3, a specific tag for epigenetic transcriptional activation, thereby acting as a corepressor. Required for de novo DNA methylation of a subset of imprinted genes during oogenesis. Acts by oxidizing the substrate by FAD to generate the corresponding imine that is subsequently hydrolyzed. Demethylates both mono- and di-methylated 'Lys-4' of histone H3. Has no effect on tri-methylated 'Lys-4', mono-, di- or tri-methylated 'Lys-9', mono-, di- or tri-methylated 'Lys-27', mono-, di- or tri-methylated 'Lys-36' of histone H3, or on mono-, di- or tri-methylated 'Lys-20' of histone H4.
  • Sequence similarities
    Belongs to the flavin monoamine oxidase family.
    Contains 1 CW-type zinc finger.
    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
  • Form
    There are 3 isoforms produced by alternative splicing.
  • Alternative names
    • amine oxidase (flavin containing) domain 1 antibody
    • amine oxidase, flavin containing 1 antibody
    • AOF1 antibody
    • bA204B7.3 antibody
    • C6orf193 antibody
    • dJ298J15.2 antibody
    • Flavin-containing amine oxidase domain-containing protein 1 antibody
    • FLJ33898 antibody
    • FLJ34109 antibody
    • FLJ43328 antibody
    • KDM1B antibody
    • KDM1B_HUMAN antibody
    • LSD2 antibody
    • lysine (K)-specific demethylase 1B antibody
    • Lysine-specific demethylase 2 antibody
    • Lysine-specific histone demethylase 1B antibody
    • Lysine-specific histone demethylase 2 antibody
    see all

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: LSD2 / AOF1 knockout HAP1 cell lysate (20 µg)
    Lane 3: PC12 cell lysate (20 µg)
    Lane 4: Raw264.7 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab193080 observed at 95 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab193080 was shown to specifically react with LSD2 / AOF1 when LSD2 / AOF1 knockout samples were used. Wild-type and LSD2 / AOF1 knockout samples were subjected to SDS-PAGE. ab193080 and ab8245 (loading control to GAPDH) were diluted at 1/2000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling LSD2 / AOF1 antibody (red) with purified ab193080 at a dilution of 1/70. Goat anti rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1/2000. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

  • All lanes : Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080) at 1/2000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 92 kDa
    Observed band size: 92 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080) at 1/2000 dilution

    Lane 1 : NIH/3T3 (Mouse embyro fibroblast cells) cell lysate
    Lane 2 : Mouse thymus lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 92 kDa
    Observed band size: 92 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080) at 1/2000 dilution

    Lane 1 : C6 (Rat glial tumor cells) cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

    Predicted band size: 92 kDa
    Observed band size: 92 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling LSD2 / AOF1 with ab193080 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on HeLa cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab193080 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma) cells labeling LSD2 / AOF1 with ab193080 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear staining on A431 cell line.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:-
    -ve control 1: ab193080 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • LSD2 / AOF1 was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate with ab193080 at 1/80 dilution.

    Lane 1: K562 whole cell lysate 10ug (Input).

    Lane 2: ab193080 IP in K562 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab193080 in K562 whole cell lysate.

    Western blot was performed from the immunoprecipitate using ab193080 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 8 seconds.

References

This product has been referenced in:
  • Pishas KI & Lessnick SL Ewing sarcoma resistance to SP-2509 is not mediated through KDM1A/LSD1 mutation. Oncotarget 9:36413-36429 (2018). Read more (PubMed: 30559927) »
See 1 Publication for this product

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