Key features and details
- Mouse monoclonal [18E8] to KDM5A / Jarid1A / RBBP2
- Suitable for: Flow Cyt, ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG2a
Product nameAnti-KDM5A / Jarid1A / RBBP2 antibody [18E8]
See all KDM5A / Jarid1A / RBBP2 primary antibodies
DescriptionMouse monoclonal [18E8] to KDM5A / Jarid1A / RBBP2
Tested applicationsSuitable for: Flow Cyt, ICC/IF, WBmore details
Species reactivityReacts with: Mouse, Human
- HeLa siRNA. HeLa RBP2 siRNA. Extracts from MCF7 cells, U2OS cells, NIH3T3 cells and J1 (mouse ES) cells.
Histone demethylase that specifically demethylates 'Lys-4' of histone H3, thereby playing a central role in histone code.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 6
Constituents: 50% Glycerol, PBS
Concentration information loading...
PurityIon Exchange Chromatography
Light chain typekappa
- Epigenetics and Nuclear Signaling
- Nuclear Signaling Pathways
- Nuclear Receptors
Our Abpromise guarantee covers the use of ab78322 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|ICC/IF||Use a concentration of 1 - 10 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Predicted molecular weight: 196 kDa.|
FunctionHistone demethylase that specifically demethylates 'Lys-4' of histone H3, thereby playing a central role in histone code. Does not demethylate histone H3 'Lys-9', H3 'Lys-27', H3 'Lys-36', H3 'Lys-79' or H4 'Lys-20'. Demethylates trimethylated and dimethylated but not monomethylated H3 'Lys-4'. May stimulate transcription mediated by nuclear receptors. May be involved in transcriptional regulation of Hox proteins during cell differentiation. May participate in transcriptional repression of cytokines such as CXCL12.
Sequence similaritiesBelongs to the JARID1 histone demethylase family.
Contains 1 ARID domain.
Contains 1 JmjC domain.
Contains 1 JmjN domain.
Contains 3 PHD-type zinc fingers.
DomainThe GSGFP motif is required for the interaction with SUZ12.
Cellular localizationNucleus > nucleolus. Occupies promoters of genes involved in RNA metabolism and mitochondrial function.
- Information by UniProt
- Histone demethylase JARID1A antibody
- JARID1A antibody
- Jumonji/ARID domain containing protein 1A antibody
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: KDM5A knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HEK293 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab78322 observed at 240 kDa. Red - loading control, ab176560, observed at 50 kDa.
ab78322 was shown to specifically react with KDM5A in wild type HAP1 cells along with additional cross reactive bands. No bands were observed when KDM5A knockout samples were examined. Ab78322 and ab176560 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 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 1/20000 dilution for 1 hour at room temperature before imaging.
ab78322 staining KDM5A / Jarid1A / RBBP2 [18E8] in HeLa cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4 % paraformaldehyde, permeabilized with Triton X-100 0.25% in PBS and blocked with 1.5% BSA for 30 minutes. Samples were incubated with primary antibody (1/200 in PBS + 1% BSA) for overnight at 4°C. An Alexa Fluor®488-conjugated Goat anti-mouse IgG polyclonal (1/1000 in PBS + 1% BSA) was used as the secondary antibody and incubated for 1 hour at room temperature. DAPI was used to stain the nuclear DNA.
All lanes : Anti-KDM5A / Jarid1A / RBBP2 antibody [18E8] (ab78322) at 1 µg/ml
Lane 1 : HeLa control siRNA
Lane 2 : HeLa RBBP2 siRNA
Lane 3 : Extracts from MCF7 cells
Lane 4 : Extracts from U2OS cells
Lane 5 : Extracts from NIH3T3 cells
Lane 6 : Extracts from J1 (mouse ES) cells
Developed using the ECL technique.
Predicted band size: 196 kDa
Observed band size: 196 kDa
Overlay histogram showing HeLa cells stained with ab78322 (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 (ab78322, 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 IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
ab78322 has been referenced in 7 publications.
- Gong F et al. Histone demethylase KDM5A regulates the ZMYND8-NuRD chromatin remodeler to promote DNA repair. J Cell Biol 216:1959-1974 (2017). PubMed: 28572115
- Maggi EC et al. Retinoblastoma-binding protein 2 (RBP2) is frequently expressed in neuroendocrine tumors and promotes the neoplastic phenotype. Oncogenesis 5:e257 (2016). PubMed: 27548814
- Penterling C et al. Depletion of Histone Demethylase Jarid1A Resulting in Histone Hyperacetylation and Radiation Sensitivity Does Not Affect DNA Double-Strand Break Repair. PLoS One 11:e0156599 (2016). WB, IF . PubMed: 27253695
- Yamamoto S et al. JARID1B is a luminal lineage-driving oncogene in breast cancer. Cancer Cell 25:762-77 (2014). WB . PubMed: 24937458
- Lopez-Bigas N et al. Genome-wide analysis of the H3K4 histone demethylase RBP2 reveals a transcriptional program controlling differentiation. Mol Cell 31:520-30 (2008). PubMed: 18722178
- Klose RJ et al. The retinoblastoma binding protein RBP2 is an H3K4 demethylase. Cell 128:889-900 (2007). PubMed: 17320163
- Christensen J et al. RBP2 belongs to a family of demethylases, specific for tri-and dimethylated lysine 4 on histone 3. Cell 128:1063-76 (2007). PubMed: 17320161