使用敲除细胞株进行验证

Anti-KDM1 / LSD1抗体[3D12] - ChIP Grade (ab90966)

概述

  • 产品名称Anti-KDM1 / LSD1抗体[3D12] - ChIP Grade
    参阅全部 KDM1 / LSD1 一抗
  • 描述
    小鼠单克隆抗体[3D12] to KDM1 / LSD1 - ChIP Grade
  • 经测试应用适用于: Flow Cyt, WB, ICC/IF, ELISA, IP, ChIPmore details
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    Recombinant fragment corresponding to Human KDM1/ LSD1.

  • 阳性对照
    • HeLa, NIH3T3, mouse P19 and human LNCaP.

性能

应用

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.

应用 Ab评论 说明
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.
ICC/IF 1/20.
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration.

靶标

  • 功能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.
  • 组织特异性Ubiquitously expressed.
  • 序列相似性Belongs to the flavin monoamine oxidase family.
    Contains 1 SWIRM domain.
  • 结构域The SWIRM domain may act as an anchor site for a histone tail.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-KDM1 / LSD1 antibody [3D12] - ChIP Grade 图像



  • Predicted band size : 93 kDa

    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 recognize KDM1 / LSD1 when KDM1 / LSD1 knockout samples were used, along with additional cross-reactive bands. 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 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

  • 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.

Anti-KDM1 / LSD1 antibody [3D12] - ChIP Grade (ab90966)参考文献

ab90966 has not yet been referenced specifically in any publications.

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