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Rabbit Recombinant Monoclonal JMJD6 antibody. Carrier free. Suitable for Flow Cyt, WB, IHC-P and reacts with Human samples.

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Images

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: 100% PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.
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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFIPFlow CytWBIHC-P
Human
Not recommended
Not recommended
Tested
Tested
Tested
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Not recommended

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Human

Dilution info

-

Notes

-

Species

Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Human, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Rat

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target data

Function

Dioxygenase that can both act as a arginine demethylase and a lysyl-hydroxylase (PubMed:24498420, PubMed:17947579, PubMed:20684070, PubMed:21060799, PubMed:22189873). Acts as a lysyl-hydroxylase that catalyzes 5-hydroxylation on specific lysine residues of target proteins such as U2AF2/U2AF65 and LUC7L2. Regulates RNA splicing by mediating 5-hydroxylation of U2AF2/U2AF65, affecting the pre-mRNA splicing activity of U2AF2/U2AF65 (PubMed:19574390). Hydroxylates its own N-terminus, which is required for homooligomerization (PubMed:22189873). In addition to peptidyl-lysine 5-dioxygenase activity, may act as an RNA hydroxylase, as suggested by its ability to bind single strand RNA (PubMed:20679243, PubMed:29176719). Also acts as an arginine demethylase which preferentially demethylates asymmetric dimethylation (PubMed:17947579, PubMed:24498420, PubMed:24360279). Demethylates histone H3 at 'Arg-2' (H3R2me) and histone H4 at 'Arg-3' (H4R3me), including mono-, symmetric di- and asymmetric dimethylated forms, thereby playing a role in histone code (PubMed:17947579, PubMed:24360279). However, histone arginine demethylation may not constitute the primary activity in vivo (PubMed:17947579, PubMed:21060799, PubMed:22189873). In collaboration with BRD4, interacts with the positive transcription elongation factor b (P-TEFb) complex in its active form to regulate polymerase II promoter-proximal pause release for transcriptional activation of a large cohort of genes. On distal enhancers, so called anti-pause enhancers, demethylates both histone H4R3me2 and the methyl cap of 7SKsnRNA leading to the dismissal of the 7SKsnRNA:HEXIM1 inhibitor complex. After removal of repressive marks, the complex BRD4:JMJD6 attract and retain the P-TEFb complex on chromatin, leading to its activation, promoter-proximal polymerase II pause release, and transcriptional activation (PubMed:24360279). Demethylates other arginine methylated-proteins such as ESR1 (PubMed:24498420). Has no histone lysine demethylase activity (PubMed:21060799). Required for differentiation of multiple organs during embryogenesis. Acts as a key regulator of hematopoietic differentiation: required for angiogenic sprouting by regulating the pre-mRNA splicing activity of U2AF2/U2AF65 (By similarity). Seems to be necessary for the regulation of macrophage cytokine responses (PubMed:15622002).

Alternative names

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Rabbit Recombinant Monoclonal JMJD6 antibody. Carrier free. Suitable for Flow Cyt, WB, IHC-P and reacts with Human samples.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

EPR23672-71

Purification technique

Affinity purification Protein A

Concentration
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Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Notes

ab282015 is the carrier-free version of ab256798.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

6 product images

  • Western blot - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (ab282015), expandable thumbnail

    Western blot - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (ab282015)

    This data was developed using ab256798, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

    Lanes 1-5: Merged signal (red and green).

    Green - ab256798 observed at 49, 147, 294kDa.

    Red - loading control ab8245 observed at 36 kDa.

    ab256798 Anti-JMJD6 antibody [EPR23672-71] was shown to react with JMJD6 in Hela cells in Western blot. Loss of signal was observed when knockout cell line

    ab266402 (JMJD6 knockout cell lysate ab257490) was used. Wild-type and JMJD6 knockout samples were subjected to SDS-PAGE.

    ab256798 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight at 1/1000 dilution and 1/20000 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 1/10000 dilution for 1 hour at room temperature before imaging.

    The molecular weight observed is consistent with what has been described in the literature (PMID:24360279).

    This antibody reacts with an unidentifiable protein around 38 kDa.

    All lanes: Western blot - Anti-JMJD6 antibody [EPR23672-71] (AB256798) at 1/1000 dilution

    Lane 1: Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

    Lane 2: JMJD6 knockout HEK-293T whole cell lysate at 20 µg

    Lane 3: A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 4: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

    Lane 5: PANC-1 (human pancreatic epithelioid carcinoma epithelial cell), whole cell lysate at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution

    Predicted band size: 46 kDa

    Observed band size: 147 kDa, 294 kDa, 49 kDa

  • Western blot - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (ab282015), expandable thumbnail

    Western blot - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (ab282015)

    This data was developed using ab256798, the same antibody clone in a different buffer formulation.

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

    Lysates were made freshly and used in WB test immediately to minimize protein degradation.

    The molecular weight observed is consistent with what has been described in the literature (PMID:24360279).

    This antibody reacts with an unidentifiable protein around 38 kDa.

    Exposure time: 48 seconds.

    All lanes: Western blot - Anti-JMJD6 antibody [EPR23672-71] (AB256798) at 1/1000 dilution

    Lane 1: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

    Lane 2: 293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051) at 1/20000 dilution

    Predicted band size: 46 kDa

    Observed band size: 147 kDa, 294 kDa, 49 kDa

  • Flow Cytometry - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (ab282015), expandable thumbnail

    Flow Cytometry - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (ab282015)

    This data was developed using ab256798, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized parental HEK293T (Human embryonic kidney epithelial cell, Right) cells / JMJD6 KO HEK293T cells (Left) labelling JMJD6 with ab256798 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Positive staining on 293T cells (ab255449), while no staining on JMJD6 knockout HEK-293T cells (ab266402).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (ab282015), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (ab282015)

    This data was developed using ab256798, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human testis tissue labelling JMJD6 with ab256798 at 1/500 dilution (1.152 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human testis. The section was incubated with ab256798 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (ab282015), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (ab282015)

    This data was developed using ab256798, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human liver tumor tissue labelling JMJD6 with ab256798 at 1/500 dilution (1.152 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human liver tumor. The section was incubated with ab256798 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (ab282015), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (ab282015)

    This data was developed using ab256798, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling JMJD6 with ab256798 at 1/500 dilution (1.152 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control: No staining on human liver. The section was incubated with ab256798 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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