Antibodies, Proteins, Kits and Reagents for Life Science

324 988 ₸ Product size

100 µl 324 988 ₸ 10 µl 130 665 ₸

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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

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

Application	Species
IHC-P	Rat Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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.
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

EPR18466
Isotype

IgG
Research areas

Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] (ab191433)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] (ab191433)

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.
Immunoprecipitation - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] (ab191433)

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.
Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] (ab191433)

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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] (ab191433)

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.
Western blot - Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] (ab191433)

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 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Anti-KDM4A / JHDM3A / JMJD2A antibody [EPR18466] (ab191433)

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