重组Anti-MLKL (phospho S358)抗体[EPR9514] (ab187091)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR9514] to MLKL (phospho S358)
- Suitable for: WB, Dot blot, IHC-P
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-MLKL (phospho S358)抗体[EPR9514]
参阅全部 MLKL 一抗 -
描述
兔单克隆抗体[EPR9514] to MLKL (phospho S358) -
宿主
Rabbit -
特异性
Stimulation may be required to allow detection of the phosphorylated protein. Please see images below for recommended treatment conditions and positive controls. -
经测试应用
适用于: WB, Dot blot, IHC-Pmore details -
种属反应性
与反应: Human
不与反应: Mouse, Rat -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab206929) -
阳性对照
- WB: HT-29 cell lysate treated with TNF alpha+ Smac mimetic+ z-VAD. HT-29 cells were treated with the indicated stimuli for 8 hours and then harvested. The final concentrations of 20 ng/ml TNF-a, 100 nM Smac mimetic, and 20 µM z-VAD were used to induce necrosis; human hepatocyte (treated with Smac/z0VAD) cell lysate. IHC-P: Human skin and melanoma tissue. Dot blot: MLKL (pS358) phospho peptide; MLKL (pT357/pS358) phospho peptide.
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常规说明
This antibody was developed through collaboration with the lab of Xiaodong Wang at the National Institute of Biological Sciences, Beijing.
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077). Or search our wide range of secondary antibodies for use with your experiment.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: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR9514 -
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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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab187091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 | (7) |
1/1000 - 1/2000. Predicted molecular weight: 54 kDa.Can be blocked with Human MLKL (phospho S358) peptide (ab206929).
We recommend using 1% SDS Hot lysis method to prepare cell lysates. For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy). |
Dot blot |
1/1000.
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IHC-P | (4) |
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Not Suitable for Mouse and Rat |
说明 |
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WB
1/1000 - 1/2000. Predicted molecular weight: 54 kDa.Can be blocked with Human MLKL (phospho S358) peptide (ab206929). We recommend using 1% SDS Hot lysis method to prepare cell lysates. For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy). |
Dot blot
1/1000. |
IHC-P
1/250 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Not Suitable for Mouse and Rat |
靶标
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序列相似性
Belongs to the protein kinase superfamily.
Contains 1 protein kinase domain. -
结构域
The protein kinase domain is predicted to be catalytically inactive. - Information by UniProt
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数据库链接
- Entrez Gene: 197259 Human
- Omim: 615153 Human
- SwissProt: Q8NB16 Human
- Unigene: 119878 Human
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别名
- 9130019I15Rik antibody
- FLJ34389 antibody
- hMLKL antibody
see all
图片
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All lanes : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 1/1000 dilution
Lane 1 : Untreated HT-29 (human colorectal adenocarcinoma) whole cell lysates 20µg
Lane 2 : HT-29 (human colorectal adenocarcinoma) treated with TNF alpha+ Smac mimetic+ z-VAD whole cell lysates 20µg
Lane 3 : HT-29 (human colorectal adenocarcinoma) treated with TNF alpha+ Smac mimetic + z-VAD and phosphatase whole cell lysates 20µg.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 54 kDa
Observed band size: 54 kDaBlocking buffer and concentration: 5% NFDM/TBST, Diluting buffer and concentration: 5% NFDM /TBST, Exposure time: 1 minute
The lysate in this image is prepared by 1%SDS Hot Lysate buffer.
For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).
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ab187091 at 1:250 staining MLKL (phospho S358) in Human melanoma tissue by immunohistochemistry (FFPE).
Antigen retrival required on FFPE tissue: HIER using 10mM Citrate buffer pH 6.0, see recommended HIER protocol
For additional IHC guideline, please see IHC resource page
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ab187091 at 1:250 staining MLKL (phospho S358) in Human skin tissue by immunohistochemistry (FFPE).
Antigen retrival required on FFPE tissue: HIER using 10mM Citrate buffer pH 6.0, see recommended HIER protocol
For additional IHC guideline, please see IHC resource page
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Dot blot analysis of MLKL peptides using ab187091 at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated secondary antibody at 1/1000 dilution. Blocking and diluting buffer was 5% NFDM/TBST, exposure time 3 minuts.
Lane 1: MLKL (pT357) phospho peptide
Lane 2: MLKL (pS358) phospho peptide
Lane 3: MLKL (pT357/pS358) phospho peptide
Lane 4: MLKL non-phospho peptide
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All lanes : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 1000 cells
Lane 1 : Untreated human hepatocyte cell lysate
Lane 2 : Treated (10 ng/ml TNF-a+100 nM Smac mimetic+20 µM z-VAD 6 h) human hepatocyte cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit HRP at 1/5000 dilution
Predicted band size: 54 kDa
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Sample: HT-29 (Human colorectal adenocarcinoma epithelial cell) treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 8 hours whole cell lysates 10 µg per lane.Lane 1 : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 0.12 µg/ml (Batch produced in 2016)
Lane 2 : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 0.17 µg/ml (Batch produced in 2015)
Lane 3 : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 0.12 µg/ml (GR212667 - batch produced in 2014)
Lane 4 : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 0.16 µg/ml (The supernatant of the clone producing ab187091)
Lane 5 : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 0.15 µg/ml (Batch produced in 2017)Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilutionBlocking and diluting buffer: 5% NFDM/TBST.
The lysate in this image is prepared by 1%SDS Hot Lysate buffer.
For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).
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All lanes : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 1/5000 dilution
Lane 1 : HT-29 (Human colorectal adenocarcinoma epithelial cell) treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 6 hr. The lysate is directly prepared by 1xSDS loading buffer.
Lane 2 : HT-29 (Human colorectal adenocarcinoma epithelial cell) treated with 20 ng/ml TNF-a, 100 nM Smac mimetic and 20 µM z-VAD for 8 hr. The lysate is prepared by 1%SDS Hot Lysate buffer method.
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: 54 kDa
Observed band size: 54 kDa
Exposure time: 3 minutesBlocking and diluting buffer: 5% NFDM/TBST.
For 1%SDS Hot Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).
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All lanes : Anti-MLKL (phospho S358) antibody [EPR9514] (ab187091) at 1/2000 dilution
Lane 1 : Untreated HT-29 lysate
Lane 2 : HT-29 cell lysate treated with TNF alpha+ Smac mimetic+ z-VAD
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 54 kDaDetails on WB tested positive control samples: HT-29 cells were treated with the indicated stimuli for 8 hr and then harvested. The final concentrations of 20 ng/ml TNF-a, 100 nM Smac mimetic, and 20 µM z-VAD were used to induce necrosis.
The lysate in this image is prepared by 1%SDS Hot Lysate buffer.
For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy).
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (191)
ab187091 被引用在 191 文献中.
- Huang D et al. Salt-inducible kinases inhibitor HG-9-91-01 targets RIPK3 kinase activity to alleviate necroptosis-mediated inflammatory injury. Cell Death Dis 13:188 (2022). PubMed: 35217652
- Jacobsen AV et al. The Lck inhibitor, AMG-47a, blocks necroptosis and implicates RIPK1 in signalling downstream of MLKL. Cell Death Dis 13:291 (2022). PubMed: 35365636
- Pagano C et al. N6-isopentenyladenosine induces cell death through necroptosis in human glioblastoma cells. Cell Death Discov 8:173 (2022). PubMed: 35393392
- Wen C et al. Concomitant pyroptotic and apoptotic cell death triggered in macrophages infected by Zika virus. PLoS One 17:e0257408 (2022). PubMed: 35446851
- 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