Recombinant Anti-KDM1/LSD1 antibody [EPR6825] - BSA and Azide free (ab224270)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6825] to KDM1/LSD1 - BSA and Azide free
- Suitable for: ChIP, Flow Cyt (Intra), ChIC/CUT&RUN-seq, ICC/IF, WB, IHC-P, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-KDM1/LSD1 antibody [EPR6825] - BSA and Azide free
See all KDM1/LSD1 primary antibodies -
Description
Rabbit monoclonal [EPR6825] to KDM1/LSD1 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ChIP, Flow Cyt (Intra), ChIC/CUT&RUN-seq, ICC/IF, WB, IHC-P, IPmore 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: HAP1, 293T, HEK293, HeLa, Jurkat, PC3, C6, Raw 264.7, PC-12, and NIH 3T3 cell lysates. IHC-P: Human testis, rat kidney, and mouse colon tissues. ICC/IF: HAP1 and HeLa cells. Flow Cyt (intra): HeLa cells. IP: Jurkat cell lysate. ChIP: HCT 116 (Human colorectal carcinoma epithelial cell). ChIC/CUT&RUN-Seq: HeLa cells.
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General notes
ab224270 is the carrier-free version of ab129195.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6825 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Anti-KDM1/LSD1 antibody [EPR6825] - Nuclear Marker and ChIP Grade (ab129195)
- Alexa Fluor® 488 Anti-KDM1/LSD1 antibody [EPR6825] - Nuclear Marker (ab184811)
- Alexa Fluor® 647 Anti-KDM1/LSD1 antibody [EPR6825] - Nuclear Marker (ab195305)
- HRP Anti-KDM1/LSD1 antibody [EPR6825] - Nuclear Marker (ab195897)
- APC Anti-KDM1 / LSD1 antibody [EPR6825] - Nuclear Marker and ChIP Grade (ab310821)
- PE Anti-KDM1 / LSD1 antibody [EPR6825] - Nuclear Marker (ab310898)
- Alexa Fluor® 594 Anti-KDM1 / LSD1 antibody [EPR6825] - Nuclear Marker (ab311646)
- Alexa Fluor® 568 Anti-KDM1 / LSD1 antibody [EPR6825] - Nuclear Marker and ChIP Grade (ab312919)
- Alexa Fluor® 555 Anti-KDM1 / LSD1 antibody [EPR6825] - Nuclear Marker and ChIP Grade (ab313131)
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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
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab224270 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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ChIP |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 110 kDa (predicted molecular weight: 92 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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Notes |
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ChIP
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 110 kDa (predicted molecular weight: 92 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
Target
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Function
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. -
Tissue specificity
Ubiquitously expressed. -
Sequence similarities
Belongs to the flavin monoamine oxidase family.
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: 23028 Human
- Entrez Gene: 99982 Mouse
- Entrez Gene: 500569 Rat
- Omim: 609132 Human
- SwissProt: O60341 Human
- SwissProt: Q6ZQ88 Mouse
- Unigene: 591518 Human
- Unigene: 28540 Mouse
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Alternative names
- Amine oxidase (flavin containing) domain 2 antibody
- Amine oxidase, flavin containing, 2 antibody
- AOF2 antibody
see all
Images
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ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/µL. 2.5X10^5 of Human wild-type HeLa cell line (ab255928) or KDM1A (LSD1) knockout HeLa cell line (ab265790) were used along with 5µg of ab129195 [EPR6825]. Assay Quality Control was conducted using 5µg Anti-CTCF(ab188408) on the same cell lines. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.
Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
This data was developed using the same antibody clone in a different buffer formulation (ab129195).
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All lanes : Anti-KDM1/LSD1 antibody [EPR6825] - Nuclear Marker and ChIP Grade (ab129195) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : KDM1A knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 92 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab129195).
Lanes 1- 2: Merged signal (red and green). Green - ab129195 observed at 110 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab129195 was shown to react with KDM1/LSD1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265790 (knockout cell lysate ab256965) was used. Wild-type HeLa and KDM1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab129195 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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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Chromatin was prepared from HCT 116 cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab129195 (red), and 20µl of protein A/G sepharose beads slurry (10µl of sepharose A beads + 10µl of sepharose G beads). 5μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129195).
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Immunocytochemistry/ Immunofluorescence - Anti-KDM1 / LSD1 antibody [EPR6825] - BSA and Azide free (ab224270)
This ICC data was generated using the same anti-KDM1/LSD! antibody clone [EPR6825] in a different buffer formulation (cat# ab129195).
ab129195 staining KDM1A/LSD1 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 ab129195 at 1μg/ml concentration and ab195889 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 Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
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All lanes : Anti-KDM1/LSD1 antibody [EPR6825] - Nuclear Marker and ChIP Grade (ab129195) at 1/10000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : KMD1 / LSD1 knockout HAP1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 92 kDaThis data was developed using the same antibody clone in a different buffer formulation (ab129195).
Lanes 1 - 4: Merged signal (red and green). Green - ab129195 observed at 110 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab129195 was shown to specifically react with KMD1 / LSD1 in wild-type HAP1 cells. No band was observed when KMD1 / LSD1 knockout samples were used. Wild-type and KMD1 / LSD1 knockout samples were subjected to SDS-PAGE. ab129195 and ab8245 (loading control to GAPDH) were both diluted 1/10,000 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 hour at room temperature before imaging. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM1 / LSD1 antibody [EPR6825] - BSA and Azide free (ab224270)Immunohistochemical staining of paraffin embedded rat kidney with purified ab129195 at a working dilution of 1/50. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129195).
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Immunocytochemistry/ Immunofluorescence - Anti-KDM1 / LSD1 antibody [EPR6825] - BSA and Azide free (ab224270)
Immunofluorescence staining of HeLa cells with purified ab129195 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab129195 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129195).
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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling KDM1/LSD1 with purified ab129195 at 1/20 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129195).
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ab129195 (purified) at 1/20 immunoprecipitating KDM1/LSD1 in 10 µg Jurkat cell lysate (Lanes 1 and 2, observed at 110 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129195).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM1 / LSD1 antibody [EPR6825] - BSA and Azide free (ab224270)Immunohistochemical staining of paraffin embedded mouse colon with purified ab129195 at a working dilution of 1/50. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129195).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM1 / LSD1 antibody [EPR6825] - BSA and Azide free (ab224270)Immunohistochemical staining of paraffin embedded human stomach carcinoma with purified ab129195 at a working dilution of 1/50. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129195).
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Immunocytochemistry/ Immunofluorescence - Anti-KDM1/LSD1 antibody [EPR6825] - BSA and Azide free (ab224270)This image is courtesy of an anonymous Abreview.
Unpurified ab129195 staining KDM1/LSD1 in human paraffin-embedded A549 lung cancer cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed using the HOPE technique and permeabilized with 0.05% Tween. Samples were incubated with primary antibody (1/100) for 45 minutes at 25°C. An Alexa Fluor®488-conjugated Donkey anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129195).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM1 / LSD1 antibody [EPR6825] - BSA and Azide free (ab224270)
Unpurified ab129195, at 1/100, staining KDM1 / LSD1 in paraffin embedded Human testis tissue by Immunohistochemistry.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab129195).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (3)
ab224270 has been referenced in 3 publications.
- Kilinc S et al. Lysine-specific demethylase-1 (LSD1) is compartmentalized at nuclear chromocenters in early post-mitotic cells of the olfactory sensory neuronal lineage. Mol Cell Neurosci 74:58-70 (2016). WB, IP, IF, IHC ; Mouse . PubMed: 26947098
- Xiong Y et al. Inhibition of Lysine-Specific Demethylase-1 (LSD1/KDM1A) Promotes the Adipogenic Differentiation of hESCs Through H3K4 Methylation. Stem Cell Rev : (2016). WB . PubMed: 27059868
- Mould AW et al. Blimp1/Prdm1 Functions in Opposition to Irf1 to Maintain Neonatal Tolerance during Postnatal Intestinal Maturation. PLoS Genet 11:e1005375 (2015). WB . PubMed: 26158850