Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] (ab191433)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18466] to KDM4A / JHDM3A / JMJD2A
- Suitable for: WB, IHC-P, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466]
See all KDM4A / JHDM3A / JMJD2A primary antibodies -
Description
Rabbit monoclonal [EPR18466] to KDM4A / JHDM3A / JMJD2A -
Host species
Rabbit -
Specificity
IHC application is recommended for human and rat. -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P RatHumanIP HumanWB MouseRatHuman -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: LNCaP, U-2 OS, HEK-293, PC-12 and NIH/3T3 whole cell lysates; human fetal kidney lysate. IHC-P: Human colon, rat colon and rat skeletal muscle tissues. IP: HeLa 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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18466 -
Isotype
IgG -
Research areas
Images
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: KDM4A / JHDM3A / JMJD2A knockout HAP1 cell lysate (20 µg)
Lane 3: U2OS cell lysate (20 µg)
Lane 4: PC12 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab191433 observed at 110 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab191433 was shown to specifically react with KDM4A / JHDM3A / JMJD2A when KDM4A / JHDM3A / JMJD2A knockout samples were used. Wild-type and KDM4A / JHDM3A / JMJD2A knockout samples were subjected to SDS-PAGE. ab191433 and ab8245 (loading control to GAPDH) were diluted 1/500 and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging. -
Immunohistochemical analysis of paraffin-embedded human colon tissue labeling KDM4A / JHDM3A / JMJD2A with ab191433 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear staining in epithelial 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.
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KDM4A / JHDM3A / JMJD2A was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab191433 at 1/80 dilution.
Western blot was performed from the immunoprecipitate using ab191433 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa whole cell lysate 10µg (Input).
Lane 2: ab191433 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab191433 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
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Lanes 1-3 : Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] (ab191433) at 1/5000 dilution
Lane 4 : Anti-MTHFD2 antibody [7] (ab191443) at 1/5000 dilution
Lane 1 : Human fetal kidney at 10 µg
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 3 : LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg
Lane 4 : U-2 OS (Human bone osteosarcoma epithelial cell line) whole call lysate at 20 µg
Secondary
Lane 1 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution
Lanes 2-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution
Developed using the ECL technique.
Predicted band size: 120 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?Exposure time: Lane 1: 1 minute; Lane 2: 3 minutes; Lane 3 and 4: 1 minute
Blocking/Dilution buffer: 5% NFDM/TBST
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Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling KDM4A / JHDM3A / JMJD2A with ab191433 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear staining in epithelial 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.
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All lanes : Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] (ab191433) at 1/5000 dilution
Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) 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
Developed using the ECL technique.
Predicted band size: 120 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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