重组Anti-RIP抗体[EPR4689-100] - BSA and Azide free (ab227843)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4689-100] to RIP - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-RIP抗体[EPR4689-100] - BSA and Azide free
参阅全部 RIP 一抗 -
描述
兔单克隆抗体[EPR4689-100] to RIP - BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WBmore details
不适用于: ICC/IF,IHC-P 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
- HeLa cells and cell lysates; Raji cell lysates.
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常规说明
ab227843 is the carrier-free version of ab178420.
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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: 7.20
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
EPR4689-100 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Conjugation kits
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Isotype control
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab227843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.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 75 kDa.
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说明 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 75 kDa. |
靶标
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功能
Essential adapter molecule for the activation of NF-kappa-B. Following different upstream signals (binding of inflammatory cytokines, stimulation of pathogen recognition receptors, or DNA damage), particular RIPK1-containing complexes are formed, initiating a limited number of cellular responses. Upon TNFA stimulation RIPK1 is recruited to a TRADD-TRAF complex initiated by TNFR1 trimerization. There, it is ubiquitinated via 'Lys-63'-link chains, inducing its association with the IKK complex, and its activation through NEMO binding of polyubiquitin chains. -
序列相似性
Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.
Contains 1 death domain.
Contains 1 protein kinase domain. -
翻译后修饰
Proteolytically cleaved by caspase-8 during TNF-induced apoptosis. Cleavage abolishes NF-kappa-B activation and enhances pro-apototic signaling through the TRADD-FADD interaction.
Autophosphorylated on serine and threonine residues.
Ubiquitinated by 'Lys-11'-, 'Lys-48'-, 'Lys-63'- and linear-linked type ubiquitin. Polyubiquitination with 'Lys-63'-linked chains by TRAF2 induces association with the IKK complex. Deubiquitination of 'Lys-63'-linked chains and polyubiquitination with 'Lys-48'-linked chains by TNFAIP3 leads to RIPK1 proteasomal degradation and consequently to the termination of the TNF- or Linear polyubiquitinated; the head-to-tail polyubiquitination is mediated by the LUBAC complex. LPS-mediated activation of NF-kappa-B. Also ubiquitinated with 'Lys-11'-linked chains. -
细胞定位
Cytoplasm. - Information by UniProt
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数据库链接
- Entrez Gene: 8737 Human
- Omim: 603453 Human
- SwissProt: Q13546 Human
- Unigene: 519842 Human
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别名
- Cell death protein RIP antibody
- FLJ39204 antibody
- OTTHUMP00000039163 antibody
see all
图片
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This Flow Cyt data was generated using the same anti-RIP antibody clone, EPR4689-100, in a different buffer formulation (cat
ab178420).
Intracellular flow cytometric analysis of permeabilized HeLa cells labeling RIP with ab178420 at 1/10 dilution (red) compared with a rabbit IgG negative control (green).
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All lanes : Anti-RIP antibody [EPR4689-100] (ab178420) at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate
Lane 2 : RIPK1 knockout THP-1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Raji cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 75 kDa
Observed band size: 76 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-RIP antibody [EPR4689-100] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab178420 was shown to bind specifically to RIP. A band was observed at 76 kDa in wild-type THP-1 cell lysates with no signal observed at this size in RIPK1 knockout cell line ab276121 (knockout cell lysate ab284210). To generate this image, wild-type and RIPK1 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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This WB data was generated using the same anti-RIP antibody clone, EPR4689-100, in a different buffer formulation (cat# ab178420).
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: RIP knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Raji cell lysate (20 µg)
Lanes 1 to 4: Merged signal (red and green). Green - ab178420 observed at 78 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab178420 was shown to specifically react with RIP when RIP knockout samples were used. Wild-type and RIP knockout samples were subjected to SDS-PAGE. ab178420 and ab8245 (loading control to GAPDH) were both diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.
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
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