Key features and details
- Mouse monoclonal [3D12] to KDM1/LSD1
- Suitable for: ICC/IF, Flow Cyt, WB
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG1
Product nameAnti-KDM1/LSD1 antibody [3D12]
See all KDM1/LSD1 primary antibodies
DescriptionMouse monoclonal [3D12] to KDM1/LSD1
Tested applicationsSuitable for: ICC/IF, Flow Cyt, WBmore details
Species reactivityReacts with: Mouse, Human
Recombinant fragment corresponding to Human KDM1/ LSD1.
- HeLa, NIH3T3, mouse P19 and human LNCaP. ICC/IF: HAP1-KDM1A cells.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferConstituent: Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
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.
|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.|
FunctionHistone 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 specificityUbiquitously expressed.
Sequence similaritiesBelongs to the flavin monoamine oxidase family.
Contains 1 SWIRM domain.
DomainThe SWIRM domain may act as an anchor site for a histone tail.
- Information by UniProt
- Amine oxidase (flavin containing) domain 2 antibody
- Amine oxidase, flavin containing, 2 antibody
- AOF2 antibody
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.
ab90966 has been referenced in 2 publications.
- Pu J et al. lncRNA MAGI2-AS3 Prevents the Development of HCC via Recruiting KDM1A and Promoting H3K4me2 Demethylation of the RACGAP1 Promoter. Mol Ther Nucleic Acids 18:351-362 (2019). PubMed: 31629962
- Zhou A et al. Nuclear GSK3ß promotes tumorigenesis by phosphorylating KDM1A and inducing its deubiquitylation by USP22. Nat Cell Biol 18:954-966 (2016). PubMed: 27501329