Anti-JMJD6 antibody (ab64575)
Key features and details
- Rabbit polyclonal to JMJD6
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Overview
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Product name
Anti-JMJD6 antibody
See all JMJD6 primary antibodies -
Description
Rabbit polyclonal to JMJD6 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide corresponding to Human JMJD6 aa 350 to the C-terminus conjugated to keyhole limpet haemocyanin.
(Peptide available asab64574) -
Positive control
- WB: THP1 and HEK-293 cell lysates; Rat Liver, Mouse Liver, Mouse Heart tissue lysates. ICC/IF: HeLa cells.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab64575 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 |
Use a concentration of 1 µg/ml.
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| WB |
Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 46 kDa).
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| Notes |
|---|
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ICC/IF
Use a concentration of 1 µg/ml. |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 46 kDa). |
Target
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Function
Dioxygenase that can both act as a histone arginine demethylase and a lysyl-hydroxylase. Acts as a lysyl-hydroxylase that catalyzes 5-hydroxylation on specific lysine residues of target proteins such as U2AF2/U2AF65 and LUC7L2. Acts as a regulator of RNA splicing by mediating 5-hydroxylation of U2AF2/U2AF65, affecting the pre-mRNA splicing activity of U2AF2/U2AF65. In addition to peptidyl-lysine 5-dioxygenase activity, may act as a RNA hydroxylase, as suggested by its ability to bind single strand RNA. Also acts as an arginine demethylase which demethylates histone H3 at 'Arg-2' (H3R2me) and histone H4 at 'Arg-3' (H4R3me), thereby playing a role in histone code. However, histone arginine demethylation may not constitute the primary activity in vivo. Has no histone lysine demethylase activity. 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. Seems to be necessary for the regulation of macrophage cytokine responses. -
Tissue specificity
Highly expressed in the heart, skeletal muscle and kidney. Expressed at moderate or low level in brain, placenta, lung, liver, pancreas, spleen, thymus, prostate, testis and ovary. Up-regulated in many patients with chronic pancreatitis. Expressed in nursing thymic epithelial cells. -
Sequence similarities
Belongs to the JMJD6 family.
Contains 1 JmjC domain. -
Domain
The nuclear localization signal motifs are necessary and sufficient to target it into the nucleus. -
Cellular localization
Nucleus > nucleoplasm. Nucleus > nucleolus. Mainly found throughout the nucleoplasm outside of regions containing heterochromatic DNA, with some localization in nucleolus. During mitosis, excluded from the nucleus and reappears in the telophase of the cell cycle. - Information by UniProt
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Database links
- Entrez Gene: 23210 Human
- Entrez Gene: 107817 Mouse
- Entrez Gene: 360665 Rat
- Omim: 604914 Human
- SwissProt: Q6NYC1 Human
- SwissProt: Q9ERI5 Mouse
- SwissProt: Q6AYK2 Rat
- Unigene: 514505 Human
see all -
Alternative names
- Apoptotic cell clearance receptor antibody
- Bifunctional arginine demethylase and lysyl-hydroxylase JMJD6 antibody
- Histone arginine demethylase JMJD6 antibody
see all
Images
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Western blot - Anti-JMJD6 antibody (ab64575)All lanes : Anti-JMJD6 antibody (ab64575) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : JMJD6 Knockout HEK293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab64575 observed at 50 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab64575 was shown to react with JMJD6 in wild-type HEK293T cells in Western blot with loss of signal observed in JMJD6 knockout cell line ab266402 (JMJD6 knockout cell lysate ab257490). Wild-type and JMJD6 knockout HEK293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab64575 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-JMJD6 antibody (ab64575)ICC/IF image of ab64575 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab64575, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 4% formaldehyde fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml. -
Western blot - Anti-JMJD6 antibody (ab64575)All lanes : Anti-JMJD6 antibody (ab64575) at 1 µg/ml
Lane 1 : THP1 cell lysate
Lane 2 : Wild-type HEK293 cell lysate
Lane 3 : JMJD6 knockout HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?Lanes 1 - 3: Merged signal (red and green). Green - ab64575 observed at 50 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab64575 was shown to react with JMJD6 in wild-type HEK-293 cells in western blot. Loss of signal was observed when JMJD6 knockout cell lysate (ab257490) was used. Wild-type and JMJD6 knockout HEK-293 cell lysates were subjected to SDS-PAGE. Membranes were blocked in non-mammalian (TBS-based) blocking solution before incubation with ab64575 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-JMJD6 antibody (ab64575)All lanes : Anti-JMJD6 antibody (ab64575) at 1 µg/ml
Lane 1 : Liver (Rat) Tissue Lysate
Lane 2 : Liver (Mouse) Tissue Lysate
Lane 3 : Heart (Mouse) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?
Additional bands at: 26 kDa, 30 kDa, 43 kDa (possible isoform). We are unsure as to the identity of these extra bands.
Exposure time: 16 minutes
Protocols
Datasheets and documents
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Datasheet download
References (7)
ab64575 has been referenced in 7 publications.
- Eddy AC et al. Differential regulation of sFlt-1 splicing by U2AF65 and JMJD6 in placental-derived and endothelial cells. Biosci Rep 40:N/A (2020). PubMed: 32039444
- Farrell A et al. Faulty oxygen sensing disrupts angiomotin function in trophoblast cell migration and predisposes to preeclampsia. JCI Insight 4:N/A (2019). PubMed: 30996134
- Wong M et al. JMJD6 is a tumorigenic factor and therapeutic target in neuroblastoma. Nat Commun 10:3319 (2019). PubMed: 31346162
- Wang L et al. Nuclear hnRNPA2B1 initiates and amplifies the innate immune response to DNA viruses. Science 365:N/A (2019). PubMed: 31320558
- Miotti S et al. Antibody-mediated blockade of JMJD6 interaction with collagen I exerts antifibrotic and antimetastatic activities. FASEB J 31:5356-5370 (2017). PubMed: 28790175
- Miller TE et al. Transcription elongation factors represent in vivo cancer dependencies in glioblastoma. Nature 547:355-359 (2017). ChIP-sequencing . PubMed: 28678782
- Tsai WC et al. Histone arginine demethylase JMJD6 is linked to stress granule assembly through demethylation of the stress granule-nucleating protein G3BP1. J Biol Chem 292:18886-18896 (2017). PubMed: 28972166
