Recombinant Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18508] to LSD2 / AOF1
- Suitable for: Flow Cyt (Intra), ICC/IF, IP, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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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 (Intra), ICC/IF, IP, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, HAP1, HCT116, 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.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18508 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
Applications
The Abpromise guarantee
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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
Purified format. |
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ICC/IF |
1/500.
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IP |
1/80.
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WB |
1/2000. Detects a band of approximately 92 kDa (predicted molecular weight: 92 kDa).
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Notes |
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Flow Cyt (Intra)
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
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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
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Database links
- Entrez Gene: 221656 Human
- Entrez Gene: 218214 Mouse
- Entrez Gene: 306819 Rat
- Omim: 613081 Human
- SwissProt: Q8NB78 Human
- SwissProt: Q8CIG3 Mouse
- Unigene: 709336 Human
- Unigene: 31259 Mouse
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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
see all
Images
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All lanes : Anti-LSD2 / AOF1 antibody [EPR18508] (ab193080) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : KDM1B knockout HeLa cell lysate
Lane 3 : HAP1 cell lysate
Lane 4 : HCT116 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 92 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab193080 observed at 95 kDa. Red - loading control ab8245 observed at 36 kDa.
ab193080 Anti-LSD2 / AOF1 antibody [EPR18508] was shown to specifically react with LSD2 / AOF1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265969 (knockout cell lysate ab258016) was used. Wild-type and LSD2 / AOF1 knockout samples were subjected to SDS-PAGE. ab193080 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Intracellular 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.
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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.
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (4)
ab193080 has been referenced in 4 publications.
- Cai S et al. Lysine-specific histone demethylase 1B (LSD2/KDM1B) represses p53 expression to promote proliferation and inhibit apoptosis in colorectal cancer through LSD2-mediated H3K4me2 demethylation. Aging (Albany NY) 12:14990-15001 (2020). PubMed: 32726297
- Patel PO et al. Investigating the role of LSD2 as an epigenetic regulator in Ewing sarcoma. Oncotarget 10:3865-3878 (2019). PubMed: 31231465
- Zhang Y et al. BAP1 links metabolic regulation of ferroptosis to tumour suppression. Nat Cell Biol 20:1181-1192 (2018). PubMed: 30202049
- Pishas KI & Lessnick SL Ewing sarcoma resistance to SP-2509 is not mediated through KDM1A/LSD1 mutation. Oncotarget 9:36413-36429 (2018). PubMed: 30559927