Anti-YTHDC1 抗体 [EPR21821-213] (ab259990)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21821-213] to YTHDC1
- Suitable for: Flow Cyt (Intra), IHC-P, WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-YTHDC1 antibody [EPR21821-213]
YTHDC1 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR21821-213] to YTHDC1 -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), IHC-P, WB, ICC/IFmore details
適用なし: IP -
種交差性
交差種: Mouse, Rat, Human
交差が予測される動物種: Cat, Dog, Pig, Zebrafish, Cynomolgus monkey, Rhesus monkey, Common marmoset, Xenopus tropicalis -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: ES-D3, HeLa, NIH/3T3, HEK-293, PC-12 lysates. IHC-P: Human testis, Mouse testis, Rat testis tissues. ICC/IF: HeLa, NIH/3T3 cells. Flow Cyt: HeLa, NIH/3T3 cells.
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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
バッファー
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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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR21821-213 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Isotype control
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab259990の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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Flow Cyt (Intra) |
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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WB |
1/1000. Predicted molecular weight: 85 kDa.
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ICC/IF |
1/50.
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特記事項 |
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Flow Cyt (Intra)
1/500. |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Predicted molecular weight: 85 kDa. |
ICC/IF
1/50. |
ターゲット情報
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機能
May be part of a signal transduction pathway that influences splice site selection. -
配列類似性
Contains 1 YTH domain. -
細胞内局在
Nucleus. Localizes to a novel subnuclear structure, the YT bodies. - Information by UniProt
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参照データベース
- Entrez Gene: 91746 Human
- Entrez Gene: 231386 Mouse
- Entrez Gene: 170956 Rat
- SwissProt: Q96MU7 Human
- SwissProt: Q9QY02 Rat
- Unigene: 175955 Human
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別名
- KIAA1966 antibody
- Putative splicing factor YT521 antibody
- splicing factor YT521-B antibody
see all
画像
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All lanes : Anti-YTHDC1 antibody [EPR21821-213] (ab259990) at 1/1000 dilution
Lane 1 : Wild-type ES-D3 (mouse embryonic pluripotent stem Cell) whole cell lysate
Lane 2 : YTHDC1 knockout ES-D3 cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : NIH/3T3 (mouse embryonic fibroblast) 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: 85 kDa
Observed band size: 100 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 1-4: Merged signal (red and green). Green - ab259990 observed at 100 kDa. Red - loading control ab8245 observed at 36 kDa.
ab259990 Anti-YTHDC1 antibody [EPR21821-213] was shown to specifically react with YTHDC1 in wild-type ES-D3 cells. Loss of signal was observed when a knockout cell line was used. Wild-type and YTHDC1 knockout samples were subjected to SDS-PAGE. ab259990 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 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 in 20000 dilution for 1 hour at room temperature before imaging.
The wild-type and YTHDC1 knockout ES-D3 lysates were kindly provided by an anonymous collaborator. -
All lanes : Anti-YTHDC1 antibody [EPR21821-213] (ab259990) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma) 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: 85 kDa
Observed band size: 100 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.
Exposure time: 15 seconds
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Immunohistochemical analysis of paraffin-embedded Human testis tissue labelling YTHDC1 with ab259990 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 testis. The section was incubated with ab220159 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 Mouse testis tissue labelling YTHDC1 with ab259990 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 mouse testis. The section was incubated with ab220159 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 Rat testis tissue labelling YTHDC1 with ab259990 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 rat testis. The section was incubated with ab220159 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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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling YTHDC1 with ab259990 at 1/50 (11.52 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing mainly nuclear staining in HeLa cells (PMID: 18669619) is observed.
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling YTHDC1 with ab259990 at 1/50 (11.52 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing mainly nuclear staining in NIH/3T3 cells (PMID: 18669619) is observed.
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling YTHDC1 with ab259990 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.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling YTHDC1 with ab259990 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.
プロトコール
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.
データシートおよび資料
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SDS download
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Datasheet download
Certificate of Compliance
参考文献 (6)
ab259990 は 6 報の論文で使用されています。
- Jiao Y et al. The m6A reader YTHDC2 promotes SIRT3 expression by reducing the stabilization of KDM5B to improve mitochondrial metabolic reprogramming in diabetic peripheral neuropathy. Acta Diabetol 60:387-399 (2023). WB ; Rat . PubMed: 36574062
- Wang D et al. YTHDC1 maintains trophoblasts function by promoting degradation of mA6-modified circMPP1. Biochem Pharmacol N/A:115456 (2023). WB ; Rat . PubMed: 36780989
- Chen J et al. YT521-B homology domain containing 1 ameliorates mitochondrial damage and ferroptosis in sleep deprivation by activating the sirtuin 1/nuclear factor erythroid-derived 2-like 2/heme oxygenase 1 pathway. Brain Res Bull 197:1-12 (2023). WB ; Mouse . PubMed: 36935054
- Jiang J et al. Identification of osteosarcoma m6A-related prognostic biomarkers using artificial intelligence: RBM15. Sci Rep 13:5255 (2023). IHC-P ; Human . PubMed: 37002245
- Zhu W et al. Vitamin D3 Suppresses Human Cytomegalovirus-Induced Vascular Endothelial Apoptosis via Rectification of Paradoxical m6A Modification of Mitochondrial Calcium Uniporter mRNA, Which Is Regulated by METTL3 and YTHDF3. Front Microbiol 13:861734 (2022). PubMed: 35359726
- Xu X et al. A positive feedback circuit between RN7SK snRNA and m6A readers is essential for tumorigenesis. Mol Ther N/A:N/A (2022). WB, ICC, RIP ; Mouse, Human . PubMed: 36566349