重组Anti-TMS1/ASC抗体[EPR23978-28] (ab283684)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23978-28] to TMS1/ASC
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-TMS1/ASC抗体[EPR23978-28]
参阅全部 TMS1/ASC 一抗 -
描述
兔单克隆抗体[EPR23978-28] to TMS1/ASC -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details
不适用于: IHC-Fr -
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: Human spleen, colon and tonsil tissue lysate; Wild-type HAP1 and THP-1 cell lysates. IHC-P: Human tonsil, colon and colon cancer tissue. ICC/IF: THP-1 cells. Flow Cyt: HeLa cells. IP: THP-1 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% 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
EPR23978-28 -
同种型
IgG -
研究领域
相关产品
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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ab283684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) |
1/5000.
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ICC/IF |
1/100.
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WB |
1/1000. Predicted molecular weight: 22 kDa.
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IP |
1/30.
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IHC-P |
1/2000. 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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Flow Cyt (Intra)
1/5000. |
ICC/IF
1/100. |
WB
1/1000. Predicted molecular weight: 22 kDa. |
IP
1/30. |
IHC-P
1/2000. 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
Promotes caspase-mediated apoptosis. This proapoptotic activity is mediated predominantly through the activation of caspase-9. May be a component of the inflammasome, a protein complex which also includes NALP2, CARD8 and CASP1 and whose function would be the activation of proinflammatory caspases. -
组织特异性
Widely expressed at low levels. Detected in peripheral blood leukocytes, lung, small intestine, spleen, thymus, colon and at lower levels in placenta, liver and kidney. Very low expression in skeletal muscle, heart and brain. Detected in the leukemia cell lines HL-60 and U937, but not in Jurkat T-cell lymphoma and Daudi Burkitt's lymphoma. Detected in the melanoma cell line WM35, but not in WM793. Not detected in HeLa cervical carcinoma cells and Molt 4 lymphocytic leukemia cells. -
序列相似性
Contains 1 CARD domain.
Contains 1 DAPIN domain. -
结构域
Interacts with CIAS1/PYPAF1 and PYDC1 via the DAPIN domain. -
翻译后修饰
Phosphorylated. -
细胞定位
Cytoplasm. Upstream of caspase activation, a redistribution from the cytoplasm to the aggregates occurs. These appear as hollow, perinuclear spherical, ball-like structures. - Information by UniProt
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数据库链接
- Entrez Gene: 29108 Human
- Omim: 606838 Human
- SwissProt: Q9ULZ3 Human
- Unigene: 499094 Human
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别名
- Apoptosis associated speck like protein containing a CARD antibody
- Apoptosis-associated speck-like protein containing a CARD antibody
- ASC antibody
see all
图片
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All lanes : Anti-TMS1/ASC antibody [EPR23978-28] (ab283684) at 1/1000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : TMS1/ASC knockout HAP1 cell lysate
Lane 3 : THP-1 (human monocytic leukemia monocyte) whole cell lysate
Lane 4 : HeLa (human cervix adenocarcinoma 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: 22 kDa
Observed band size: 17,24 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 3% NFDM/TBST
Negative control: HeLa (PMID:11103776;10567338).
Lanes 1-4: Merged signal (red and green). Green - ab283684 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.
Ab283684 Anti-TMS1/ASC antibody [EPR23978-28] was shown to specifically react with TMS1/ASC in wild-type HAP1 cells. Loss of signal was observed when knockout cell line (knockout cell lysate) was used. Wild-type and TMS1/ASC knockout samples were subjected to SDS-PAGE. ab283684 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.
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All lanes : Anti-TMS1/ASC antibody [EPR23978-28] (ab283684) at 1/1000 dilution
Lane 1 : Human spleen tissue lysate
Lane 2 : Human colon tissue lysate
Lane 3 : Human tonsil tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 22 kDa
Observed band size: 12,17,24 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
ASC has four isoforms. 24-kDa ASC, 17-kDa ASC-b and 12-kDa ASC-C are observed. The molecular weight observed is consistent with what has been described in the literature (PMID:20482797).
Exposure time: 81 seconds
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Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling TMS1/ASC with ab283684 at 1/2000 (0.247 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human tonsil. The section was incubated with ab283684 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
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 colon tissue labelling TMS1/ASC with ab283684 at 1/2000 (0.247 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon. The section was incubated with ab283684 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
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 colon cancer tissue labelling TMS1/ASC with ab283684 at 1/2000 (0.247 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon cancer (PMID: 14643031). The section was incubated with ab283684 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 LeicaDS9800 (Bond™ Polymer Refine Detection).
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 THP-1 cells labelling TMS1/ASC with ab283684 at 1/100 (4.94 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic taining in THP-1 cell line. Negative control: HeLa (PMID: 11103776). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell, Left) / THP-1 (Human monocytic leukemia monocyte, Right) cells labelling TMS1/ASC with ab283684 at 1/5000 dilution (0.01ug) (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, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: HeLa (PMID: 11103776).
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TMS1/ASC was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte) whole cell lysate 20 ug with ab283684 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283684 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: THP-1 (human monocytic leukemia monocyte) whole cell lysate 20 ug
Lane 2: ab283684 IP in THP-1 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab283684 in THP-1 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds
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 (3)
ab283684 被引用在 3 文献中.
- Mo R et al. Nicotine promotes chronic obstructive pulmonary disease via inducing pyroptosis activation in bronchial epithelial cells. Mol Med Rep 25:N/A (2022). PubMed: 35059736
- Su JQ et al. The Inflammasomes Adaptor Protein PYCARD Is a Potential Pyroptosis Biomarker Related to Immune Response and Prognosis in Clear Cell Renal Cell Carcinoma. Cancers (Basel) 14:N/A (2022). PubMed: 36291776
- Jiang A et al. Establishment of a prognosis Prediction Model Based on Pyroptosis-Related Signatures Associated With the Immune Microenvironment and Molecular Heterogeneity in Clear Cell Renal Cell Carcinoma. Front Oncol 11:755212 (2021). PubMed: 34804944