Recombinant Anti-KDM2A antibody [EPR18602] KO Tested (ab191387)

Product name

Anti-KDM2A antibody [EPR18602]
See all KDM2A primary antibodies
Description

Rabbit monoclonal [EPR18602] to KDM2A
Host species

Rabbit
Tested applications

Suitable for: ICC/IF, IP, WB, IHC-Pmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human brain lysate; HeLa, Jurkat, K562, C6, PC-12, NIH/3T3 and RAW 264.7 whole cell lysates; rat brain lysate; mouse testis and brain lysates. IHC-P: Human, mouse and rat colon tissues. ICC/IF: HeLa and Jurkat cells. IP: Mouse brain whole cell lysate.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18602
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human KDM2A protein (ab151923)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab191387 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
ICC/IF		1/1000.
IP		1/80.
WB		1/1000. Detects a band of approximately 133, 90 kDa (predicted molecular weight: 133 kDa).
IHC-P		1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Notes
ICC/IF 1/1000.
IP 1/80.
WB 1/1000. Detects a band of approximately 133, 90 kDa (predicted molecular weight: 133 kDa).
IHC-P 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Function

Histone demethylase that specifically demethylates 'Lys-36' of histone H3, thereby playing a central role in histone code. Preferentially demethylates dimethylated H3 'Lys-36' residue while it has weak or no activity for mono- and tri-methylated H3 'Lys-36'. May also recognize and bind to some phosphorylated proteins and promote their ubiquitination and degradation. Required to maintain the heterochromatic state. Associates with centromeres and represses transcription of small non-coding RNAs that are encoded by the clusters of satellite repeats at the centromere. Required to sustain centromeric integrity and genomic stability, particularly during mitosis.
Tissue specificity

Widely expressed, with highest levels in brain, testis and ovary, followed by lung.
Sequence similarities

Belongs to the JHDM1 histone demethylase family.
Contains 1 CXXC-type zinc finger.
Contains 1 F-box domain.
Contains 1 JmjC domain.
Contains 6 LRR (leucine-rich) repeats.
Contains 1 PHD-type zinc finger.
Domain

The JmjC domain mediates demethylation activity and is required for satellite silencing.
The CXXC zinc finger preferentially recognizes nonmethylated CpG DNA, and binding is blocked when the CpG DNA is methylated.
Post-translational
modifications

Phosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localization

Nucleus > nucleoplasm. Punctate expression throughout the nucleoplasm and enriched in the perinucleolar region. Specifically nucleates at CpG islands where it's presence results in chromatin depleted in H3K36me2.
Target information above from: UniProt accession Q9Y2K7 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 22992 Human
- Entrez Gene: 225876 Mouse
- Entrez Gene: 361700 Rat
- Omim: 605657 Human
- SwissProt: Q9Y2K7 Human
- SwissProt: P59997 Mouse
- Unigene: 124147 Human
- Unigene: 31941 Mouse
Alternative names
- [Histone-H3]-lysine-36 demethylase 1A antibody
- CXXC-type zinc finger protein 8 antibody
- CXXC8 antibody
- F box / LRR repeat protein 11 antibody
- F box and leucine rich repeat protein 11 antibody
- F box protein FBL7 antibody
- F-box and leucine-rich repeat protein 11 antibody
- F-box protein FBL7 antibody
- F-box protein Lilina antibody
- F-box/LRR-repeat protein 11 antibody
- FBL11 antibody
- FBL7 antibody
- FBXL11 antibody
- JHDM1A antibody
- JmjC domain-containing histone demethylation protein 1A antibody
- Jumonji C domain containing histone demethylase 1A antibody
- kdm2a antibody
- KDM2A_HUMAN antibody
- LILINA antibody
- Lysine (K) specific demethylase 2A antibody
- Lysine-specific demethylase 2A antibody
see all

Western blot - Anti-KDM2A antibody [EPR18602] (ab191387)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: KDM2A knockout HAP1 whole cell lysate (20 µg)
Lane 3: Hela whole cell lysate (20 µg)
Lane 4: Jurkat whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab191387 observed at 133 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab191387 was shown to specifically react with KDM2A in wild-type HAP1 cells. No band was observed when KDM2A knockout cells were examined. Wild-type and KDM2A knockout samples were subjected to SDS-PAGE. Ab191387 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1.313 ug/ml and 1/20,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-KDM2A antibody [EPR18602] (ab191387)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling KDM2A with ab191387 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (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 Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows:-
-ve control 1: ab191387 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM2A antibody [EPR18602] (ab191387)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling KDM2A with ab191387 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on epithelium cells of human colon is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-KDM2A antibody [EPR18602] (ab191387)

Anti-KDM2A antibody [EPR18602] (ab191387) at 1/5000 dilution + Human brain lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 133 kDa
Observed band size: 133 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-KDM2A antibody [EPR18602] (ab191387)

All lanes : Anti-KDM2A antibody [EPR18602] (ab191387) at 1/1000 dilution

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 133 kDa
Observed band size: 133,90 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1 and 2: 8 seconds; Lane 3: 15 seconds.
This antibody recognizes two isoforms. The predicted MW are 133kDa and 90kDa respectively.
Western blot - Anti-KDM2A antibody [EPR18602] (ab191387)

All lanes : Anti-KDM2A antibody [EPR18602] (ab191387) at 1/1000 dilution

Lane 1 : Rat brain lysate
Lane 2 : Mouse testis lysate
Lane 3 : Mouse brain lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 133 kDa
Observed band size: 133 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1 and 3: 3 minutes; Lane 2: 30 seconds.
Western blot - Anti-KDM2A antibody [EPR18602] (ab191387)

All lanes : Anti-KDM2A antibody [EPR18602] (ab191387) at 1/1000 dilution

Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 4 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 133 kDa
Observed band size: 133 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1 and 2: 15 seconds; Lane 3: 8 seconds; Lane 4: 30 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM2A antibody [EPR18602] (ab191387)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling KDM2A with ab191387 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on epithelium cells of mouse colon is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM2A antibody [EPR18602] (ab191387)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling KDM2A with ab191387 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on epithelium cells of rat colon is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-KDM2A antibody [EPR18602] (ab191387)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling KDM2A with ab191387 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on Jurkat cell line. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows:-
-ve control 1: ab191387 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Immunoprecipitation - Anti-KDM2A antibody [EPR18602] (ab191387)

KDM2A was immunoprecipitated from 1mg of mouse brain whole cell lysate with ab191387 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab191387 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: Mouse brain whole cell lysate 10µg (Input).

Lane 2: ab191387 IP in Mouse brain whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab191387 in Mouse brain whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 30 seconds.
Anti-KDM2A antibody [EPR18602] (ab191387)

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab191387? Please let us know so that we can cite the reference in this datasheet.

ab191387 has been referenced in 5 publications.

Shou T et al. MicroRNA-3666 suppresses the growth and migration of glioblastoma cells by targeting KDM2A. Mol Med Rep 19:1049-1055 (2019). PubMed: 30483744
Lu DH et al. Lysine demethylase 2A promotes the progression of ovarian cancer by regulating the PI3K pathway and reversing epithelial-mesenchymal transition. Oncol Rep 41:917-927 (2019). PubMed: 30483796
Xu WH et al. Knockdown of KDM2A inhibits proliferation associated with TGF-ß expression in HEK293T cell. Mol Cell Biochem N/A:N/A (2019). PubMed: 30604066
Ou R et al. HPV16 E7-induced upregulation of KDM2A promotes cervical cancer progression by regulating miR-132-radixin pathway. J Cell Physiol 234:2659-2671 (2019). PubMed: 30132864
Fu E et al. Histone demethylase Kdm2a regulates germ cell genes and endogenous retroviruses in embryonic stem cells. Epigenomics 11:751-766 (2019). PubMed: 31172793

Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (5)

Customer reviews and Q&As

Post-translational
modifications