重组Anti-DFNA5/GSDME抗体[EPR19859] - N-terminal (ab215191)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19859] to DFNA5/GSDME - N-terminal
- Suitable for: WB, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-DFNA5/GSDME抗体[EPR19859] - N-terminal
参阅全部 DFNA5/GSDME 一抗 -
描述
兔单克隆抗体[EPR19859] to DFNA5/GSDME - N-terminal -
宿主
Rabbit -
特异性
Our previous testing data indicates that this antibody detects no signal or very weak signal in mouse heart, spleen, lung, stomach, testicle, skin, pancreas, muscle and lymph node tissues. Final results may be affected by the expression level of the tested samples. -
经测试应用
适用于: WB, IP, Flow Cyt (Intra)more details -
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: Mouse brain lysate; SH-SY5Y and EMT6 whole cell lysates; Human fetal brain lysate; Rat brain lysates; HEK-293 transfected with DDDDK tagged DFNA5/GSDME (N-terminal) expression vector whole cell lysate; SGC-7901 cell lysate. Flow Cyt (intra).: SH-SY5Y and EMT6 cells. IP: EMT6 whole cell lysate
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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 long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol, PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR19859 -
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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Conjugation kits
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Isotype control
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab215191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 | (3) |
1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
Our previous testing data indicates that this antibody detects no signal or very weak signal in mouse heart, spleen, lung, stomach, testicle, skin, pancreas, muscle and lymph node tissues. Final results may be affected by the expression level of the tested samples. |
IP |
1/30.
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Flow Cyt (Intra) |
1/60.
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说明 |
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WB
1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa). Our previous testing data indicates that this antibody detects no signal or very weak signal in mouse heart, spleen, lung, stomach, testicle, skin, pancreas, muscle and lymph node tissues. Final results may be affected by the expression level of the tested samples. |
IP
1/30. |
Flow Cyt (Intra)
1/60. |
靶标
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组织特异性
Expressed in cochlea. Low level of expression in heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas, with highest expression in placenta. -
疾病相关
Defects in DFNA5 are the cause of deafness autosomal dominant type 5 (DFNA5) [MIM:600994]. DFNA5 is a form of sensorineural hearing loss. Sensorineural deafness results from damage to the neural receptors of the inner ear, the nerve pathways to the brain, or the area of the brain that receives sound information. -
序列相似性
Belongs to the gasdermin family. - Information by UniProt
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数据库链接
- Entrez Gene: 1687 Human
- Entrez Gene: 54722 Mouse
- Entrez Gene: 353316 Rat
- Omim: 608798 Human
- SwissProt: O60443 Human
- SwissProt: Q9Z2D3 Mouse
- Unigene: 520708 Human
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别名
- 2310037D07Rik antibody
- 4932441K13Rik antibody
- Deafness, autosomal dominant 5 antibody
see all
图片
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All lanes : Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 1/1000 dilution
Lane 1 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 2 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDaExposure times:
Lane 1: 15s
Lane 2: 3 min
The band around 35kDa is GSDME-NT which might be cleaved by activated caspase-3 (PMID: 33589596, PMID: 31347748,PMID: 35623158).
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized EMT6 (mouse mammary carcinoma cell line) cell line labeling DFNA5/GSDMEwith ab215191 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor®488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
EMT6 cells were kindly provided by our collaborator Dr. Feng Shao, NIBS
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ab215191 at 1/30 immunoprecipitating DFNA5/GSDME in EMT6 (mouse mammary carcinoma cell line) whole cell lysate.
Lane 1 (input): EMT6 whole cell lysate (10μg)
Lane 2 (+): ab215191 + EMT6 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab215191 in EMT6 whole cell lysate
For western blotting, ab215191 at 1/500. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
EMT6 cells were kindly provided by our collaborator Dr. Feng Shao, NIBS
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All lanes : Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 0.7 µg/ml
Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse kidney tissue lysate
Lane 3 : Mouse liver tissue lysate
Lane 4 : Mouse thymus tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG) at 1/2000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDa
Exposure time: 3 minutesBlocking and diluting buffer: 5% NFDM/TBST
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All lanes : Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 1/1000 dilution
Lane 1 : Wild Type SGC-7901 (human papillomavirus-related endocervical adenocarcinoma cell line) cell lysates
Lane 2 : DFNA5/GSDME Knockout SGC-7901 cell lysates
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/2000 dilution
Predicted band size: 55 kDahttps://www.sciencedirect.com/journal/biochemical-and-biophysical-research-communications
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized SH-SY5Y (human neuroblastoma cell line from bone marrow) cell line labeling DFNA5/GSDME with ab215191 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
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Different batches of ab215191 were tested on SH-SY5Y (Human neuroblastoma epithelial cell) lysate at 0.6 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 55 kDa.
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All lanes : Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 1/5000 dilution
Lane 1 : SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate
Lane 2 : DFNA5/GSDME knockout SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 55 kDa
Observed band size: 55 kDa
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The cell lysates were kindly provided by our collaborator, Dr. Feng Shao, NIBS.
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All lanes : Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 1/10000 dilution
Lane 1 : HEK-293 (human epithelial cell line from embryonic kidney) transfected with a control vector containing DDDDK tag, whole cell lysate
Lane 2 : HEK-293 (human epithelial cell line from embryonic kidney) transfected with DDDDK tagged DFNA5/GSDME (N-terminal) expression vector, whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 55 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
This antibody is specific to the N-terminus of DFNA5/GSDME.
The cell lysates were kindly provided by our collaborator, Dr. Feng Shao, NIBS.
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Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 1/1000 dilution + Human fetal brain lysate at 10 µg
Secondary
VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 55 kDa
Observed band size: 55 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 28459430).
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All lanes : Anti-DFNA5/GSDME antibody [EPR19859] - N-terminal (ab215191) at 1/1000 dilution
Lane 1 : EMT6 (mouse mammary carcinoma cell line) whole cell lysate
Lane 2 : Mouse brain lysate
Lane 3 : Rat brain lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 55 kDa
Observed band size: 55 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 28459430).
The EMT6 cell lysate was kindly provided by our collaborator Dr. Feng Shao, NIBS.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (97)
ab215191 被引用在 97 文献中.
- Xia B et al. Extracellular vesicles mediate antibody-resistant transmission of SARS-CoV-2. Cell Discov 9:2 (2023). PubMed: 36609376
- Li F et al. GSDME Increases Chemotherapeutic Drug Sensitivity by Inducing Pyroptosis in Retinoblastoma Cells. Oxid Med Cell Longev 2022:2371807 (2022). PubMed: 35480866
- Xia J et al. Mitochondrial Protein UCP1 Inhibits the Malignant Behaviors of Triple-negative Breast Cancer through Activation of Mitophagy and Pyroptosis. Int J Biol Sci 18:2949-2961 (2022). PubMed: 35541900
- Ma L et al. A novel 3-acyl isoquinolin-1(2H)-one induces G2 phase arrest, apoptosis and GSDME-dependent pyroptosis in breast cancer. PLoS One 17:e0268060 (2022). PubMed: 35551332
- Gu J et al. Campylobacter jejuni Cytolethal Distending Toxin Induces GSDME-Dependent Pyroptosis in Colonic Epithelial Cells. Front Cell Infect Microbiol 12:853204 (2022). PubMed: 35573789