重组Anti-YTHDC2抗体[EPR21820-49] - BSA and Azide free (ab259277)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21820-49] to YTHDC2 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), IHC-P, IP, WB, IHC-Fr
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-YTHDC2抗体[EPR21820-49] - BSA and Azide free
参阅全部 YTHDC2 一抗 -
描述
兔单克隆抗体[EPR21820-49] to YTHDC2 - BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), IHC-P, IP, WB, IHC-Frmore details
不适用于: ICC/IF -
种属反应性
与反应: Mouse, Rat, 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 mESC, HT-1080, HeLa, HEK-293, NIH/3T3, PC-12 whole cell lysates. IHC-P: Human testis, Mouse testis and Rat testis tissues. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate. IHC-Fr: Mouse testis, Rat testis tissues.
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常规说明
ab259277 is the carrier-free version of ab220160.
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.
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. Do Not Freeze. -
存储溶液
pH: 7.2
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR21820-49 -
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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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab259277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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 160 kDa (predicted molecular weight: 160 kDa).
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IHC-Fr |
Use at an assay dependent concentration.
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说明 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 160 kDa (predicted molecular weight: 160 kDa). |
IHC-Fr
Use at an assay dependent concentration. |
靶标
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序列相似性
Belongs to the DEAD box helicase family. DEAH subfamily.
Contains 2 ANK repeats.
Contains 1 helicase ATP-binding domain.
Contains 1 helicase C-terminal domain.
Contains 1 R3H domain.
Contains 1 YTH domain. - Information by UniProt
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数据库链接
- Entrez Gene: 64848 Human
- Entrez Gene: 240255 Mouse
- Entrez Gene: 307446 Rat
- SwissProt: Q9H6S0 Human
- SwissProt: B2RR83 Mouse
- Unigene: 231942 Human
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别名
- Probable ATP-dependent RNA helicase YTHDC2 antibody
- YTDC2_HUMAN antibody
- YTH domain containing 2 antibody
- Ythdc2 antibody
图片
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All lanes : Anti-YTHDC2 antibody [EPR21820-49] (ab220160) at 1/1000 dilution
Lane 1 : Wild-type mESC (mouse embryo stem cell) whole cell lysate
Lane 2 : YTHDC2 knockout mESC whole cell lysate
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 160 kDa
Observed band size: 160 kDa
Exposure time: 3 minutesBlocking and dilution bufer: 5% NFDM/TBST.
The wild-type and YTHDC2 knockout cell lysates were kindly provided by an anonymous collaborator.
ab220160 was shown to specifically react with YTHDC2 in wild-type mESC cells as signal was lost in YTHDC2 knockout cells. Wild-type and YTHDC2 knockout samples were subjected to SDS-PAGE. ab220160 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220160).
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YTHDC2 was immunoprecipitated from 0.35 mg of HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab220160 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220160 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.
Lane 1: HeLa whole cell lysate 10 μ (Input).
Lane 2: ab220160IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220160 in HeLa whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220160).
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Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling YTHDC2 with ab220160 at 1/500 dilution (1.19 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on rat testis (PMID:28380054) Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220160).
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Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling YTHDC2 with ab220160 at 1/500 dilution (1.19 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on mouse testis (PMID:28380054) Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220160).
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Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling YTHDC2 with ab220160 at 1/500 dilution (1.19 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on human testis (PMID:28380054) Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220160).
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling YTHDC2 with ab220160 at 1/600 (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220160).
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220160).
Immunohistochemical analysis of 4% paraformaldehyde fixed, 0.2% Triton X-100 permeabilised mouse testis tissue labeling YTHDC2 with ab220160 at 1/50 dilution (11 ug/mL). Followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (2 ug/mL). Showing cytoplasmic staining on mouse testis. Nuclear counterstain: DAPI.
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220160).
Negative control: (PMID: 29087293)
Immunohistochemical analysis of 4% paraformaldehyde fixed, 0.2% Triton X-100 permeabilised mouse heart tissue labeling YTHDC2 with ab220160 at 1/50 dilution (11 ug/mL). Followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (2 ug/mL). No staining observed on mouse heart. Nuclear counterstain: DAPI.
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220160).
Immunohistochemical analysis of 4% paraformaldehyde fixed, 0.2% Triton X-100 permeabilised rat testis tissue labeling YTHDC2 with ab220160 at 1/50 dilution (11 ug/mL). Followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (2 ug/mL). Showing cytoplasmic staining on rat testis. Nuclear counterstain: DAPI.
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab220160).
Negative control: (PMID: 29087293)
Immunohistochemical analysis of 4% paraformaldehyde fixed, 0.2% Triton X-100 permeabilised rat heart tissue labeling YTHDC2 with ab220160 at 1/50 dilution (11 ug/mL). Followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (2 ug/mL). No staining observed on rat heart. Nuclear counterstain: DAPI.
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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Datasheet download
Certificate of Compliance
文献 (1)
ab259277 被引用在 1 文献中.
- Wang X et al. Long noncoding RNA GAS5-AS1 suppresses growth and metastasis of cervical cancer by increasing GAS5 stability. Am J Transl Res 11:4909-4921 (2019). PubMed: 31497208