重组Anti-MLKL抗体[EPR17514] (ab184718)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17514] to MLKL
- Suitable for: ICC/IF, WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-MLKL抗体[EPR17514]
参阅全部 MLKL 一抗 -
描述
兔单克隆抗体[EPR17514] to MLKL -
宿主
Rabbit -
经测试应用
适用于: ICC/IF, WB, IHC-Pmore details -
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: HUVEC, HT-29 and HeLa whole cell lysates; Human fetal kidney lysate. IHC-P: Human tonsil and colonic adenocarcinoma tissues.
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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: 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
EPR17514 -
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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KO cell lines
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KO cell lysates
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab184718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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ICC/IF | (1) |
1/200.
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WB | (4) |
1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
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IHC-P |
1/400. 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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ICC/IF
1/200. |
WB
1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa). |
IHC-P
1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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 antibody [EPR17514] (ab184718) at 1/1000 dilution
Lane 1 : HUVEC cell lysate
Lane 2 : HT-29 cell lysate
Lane 3 : Wild-type HeLa cell lysate
Lane 4 : MLKL knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 54 kDaLanes 1 - 4: Merged signal (red and green). Green - ab184718 observed at 54 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab184718 was shown to react with MLKL in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255408 (knockout cell lysate ab263788) was used. Wild-type and MLKL knockout samples were subjected to SDS-PAGE. ab184718 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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-MLKL antibody [EPR17514] (ab184718) at 1/1000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : MLKL knockout HAP1 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Huvec cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 54 kDaLanes 1 - 4: Merged signal (red and green). Green - ab184718 observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab184718 was shown to recognize MLKL in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when MLKL knockout samples were examined. Wild-type and MLKL knockout samples were subjected to SDS-PAGE. ab184718 and ab8245 (loading control to GAPDH) were diluted to 1/1000 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10,000 dilution for 1 hour at room temperature before imaging. -
Ab184718 staining MLKL in HT-29 (Human colorectal adenocarcinoma epithelial cell) cells by Immunocytochemistry (ICC). Cells were fixed with 100% Methanol. Samples were incubated with primary antibody at 1/200 dilution (6.5μg/ml). An AlexaFluor® 488 Goat anti-Rabbit (ab150077) was used as the secondary antibody at 1/1000 dilution (2μg/ml). Ab195889 , Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used as the counterstain antibody (1/200 dilution, 2.5 μg/ml . DAPI was used as a nuclear counterstain. Confocal image showing cytoplasmic staining on HT-29 cell line.
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Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling MLKL with ab184718 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Cytoplasmic staining on the lymphocytes of human tonsil is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human colonic adenocarcinoma tissue labeling MLKL with ab184718 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (ab97051) at 1/500 dilution. Cytoplasmic staining on tumor cells of human colonic adenocarcinoma is observed. Counter stained with Hematoxylin.
Negative control: Using PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-MLKL antibody [EPR17514] (ab184718) at 1/20000 dilution
Lane 1 : HUVEC (Human umbilical vein endothelial cell line) whole cell lysate
Lane 2 : HT-29 (Human colorectal adenocarcinoma cells) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 54 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-MLKL antibody [EPR17514] (ab184718) at 1/1000 dilution + Human fetal kidney lysate at 10 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 54 kDa
Observed band size: 54 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (64)
ab184718 被引用在 64 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
- Oka K et al. Resistance to chemical carcinogenesis induction via a dampened inflammatory response in naked mole-rats. Commun Biol 5:287 (2022). PubMed: 35354912
- Ellegast JM et al. Unleashing Cell-Intrinsic Inflammation as a Strategy to Kill AML Blasts. Cancer Discov 12:1760-1781 (2022). PubMed: 35405016
- 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
- Lee AR et al. Mitochondrial Transplantation Ameliorates the Development and Progression of Osteoarthritis. Immune Netw 22:e14 (2022). PubMed: 35573148