The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 110 kDa (predicted molecular weight: 93 kDa).
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.
Belongs to the flavin monoamine oxidase family. Contains 1 SWIRM domain.
The SWIRM domain may act as an anchor site for a histone tail.
FAD binding protein BRAF35 HDAC complex, 110 kDa subunit antibody
Flavin-containing amine oxidase domain-containing protein 2 antibody
KDM 1 antibody
LSD 1 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
Western blot - Anti-KDM1 / LSD1 antibody [LSD1-12] (ab53269)
Predicted band size : 93 kDa
Lane 1: Wild-type HAP1 whole cell lysate (40 µg) Lane 2: KDM1A knockout HAP1 whole cell lysate (40 µg) Lane 3: HeLa whole cell lysate (40 µg) Lane 4: Jurkat whole cell lysate (40 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab53269 observed at 92 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab53269 was shown to specifically react with KDM1 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when KDM1 knockout samples were used. Wild-type and KDM1 knockout samples were subjected to SDS-PAGE. Ab53269 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 2 µg/ml concentration and 1/10,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.
ab53269 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 ab53269 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.