重组Anti-JMJD6抗体[EPR23672-71] (ab256798)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23672-71] to JMJD6
- Suitable for: WB, Flow Cyt, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-JMJD6抗体[EPR23672-71]
参阅全部 JMJD6 一抗 -
描述
兔单克隆抗体[EPR23672-71] to JMJD6 -
宿主
Rabbit -
经测试应用
适用于: WB, Flow Cyt, IHC-Pmore details
不适用于: ICC/IF or IP -
种属反应性
与反应: Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Wild-type HEK-293T, A549, HeLa, PANC-1, 293T lysates. IHC-P: Human testis, Human liver tumor tissues. Flow Cyt: HEK293T cells.
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常规说明
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.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR23672-71 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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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
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab256798于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB |
1/1000. Predicted molecular weight: 46 kDa.
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Flow Cyt |
1/500.
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IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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说明 |
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WB
1/1000. Predicted molecular weight: 46 kDa. |
Flow Cyt
1/500. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
靶标
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功能
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. -
组织特异性
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. -
序列相似性
Belongs to the JMJD6 family.
Contains 1 JmjC domain. -
结构域
The nuclear localization signal motifs are necessary and sufficient to target it into the nucleus. -
细胞定位
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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数据库链接
- Entrez Gene: 23210 Human
- Omim: 604914 Human
- SwissProt: Q6NYC1 Human
- Unigene: 514505 Human
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别名
- Apoptotic cell clearance receptor antibody
- Bifunctional arginine demethylase and lysyl-hydroxylase JMJD6 antibody
- Histone arginine demethylase JMJD6 antibody
see all
图片
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All lanes : Anti-JMJD6 antibody [EPR23672-71] (ab256798) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 2 : JMJD6 knockout HEK-293T whole cell lysate
Lane 3 : A549 (human lung carcinoma epithelial cell) whole cell lysate
Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 5 : PANC-1 (human pancreatic epithelioid carcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
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: 147,294,49 kDa why is the actual band size different from the predicted?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.
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All lanes : Anti-JMJD6 antibody [EPR23672-71] (ab256798) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : 293T (human embryonic kidney epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 46 kDa
Observed band size: 147,294,49 kDa why is the actual band size different from the predicted?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
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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.
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Immunohistochemical analysis of paraffin-embedded Human liver tumor tissue labelling JMJD6 with ab256798 at 1/500 (1.152 ug/ml) dilution 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
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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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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).
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (0)
ab256798 尚未被引用在任何文献中。