重组Anti-Cytokeratin 18抗体[E431-1] (ab32118)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E431-1] to Cytokeratin 18
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Cytokeratin 18抗体[E431-1]
参阅全部 Cytokeratin 18 一抗 -
描述
兔单克隆抗体[E431-1] to Cytokeratin 18 -
宿主
Rabbit -
特异性
Human Cytokeratin 18 (K18) was used as immunogen after isolation from cells pre-treated with okadaic acid or pervanadate to promote Tyr hyperphosphorylation. -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
种属反应性
与反应: Human -
免疫原
Full length native protein (purified) corresponding to Human Cytokeratin 18.
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阳性对照
- WB: A431 cell lysate. IHC-P: Human gastric adenocarcinoma, kidney, colon, breast carcinoma, brain, stomach and glioma tissue. ICC/IF: HT-29 cells; HeLa cells; (negative: U87-MG cells). Flow Cyt (intra): MCF7 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: 7.20
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
E431-1 -
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
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32118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) |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/2000. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).
For unpurified use at 1/10000 |
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IHC-P | (3) |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/100 - 1/500.
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说明 |
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Flow Cyt (Intra)
1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/2000. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa). For unpurified use at 1/10000 |
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/500. |
靶标
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功能
Involved in the uptake of thrombin-antithrombin complexes by hepatic cells (By similarity). When phosphorylated, plays a role in filament reorganization. Involved in the delivery of mutated CFTR to the plasma membrane. Together with KRT8, is involved in interleukin-6 (IL-6)-mediated barrier protection. -
组织特异性
Expressed in colon, placenta, liver and very weakly in exocervix. Increased expression observed in lymph nodes of breast carcinoma. -
疾病相关
Defects in KRT18 are a cause of cirrhosis (CIRRH) [MIM:215600]. -
序列相似性
Belongs to the intermediate filament family. -
翻译后修饰
Phosphorylation at Ser-34 increases during mitosis. Hyperphosphorylated at Ser-53 in diseased cirrhosis liver. Phosphorylation increases by IL-6.
Proteolytically cleaved by caspases during epithelial cell apoptosis. Cleavage occurs at Asp-238 by either caspase-3, caspase-6 or caspase-7.
O-glycosylated at multiple sites; glycans consist of single N-acetylglucosamine residues. -
细胞定位
Cytoplasm > perinuclear region. - Information by UniProt
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数据库链接
- Entrez Gene: 3875 Human
- Omim: 148070 Human
- SwissProt: P05783 Human
- Unigene: 406013 Human
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别名
- Cell proliferation inducing gene 46 protein antibody
- Cell proliferation inducing protein 46 antibody
- Cell proliferation-inducing gene 46 protein antibody
see all
图片
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Flow cytometry overlay histogram showing left MCF7 positive cells and right negative A375 stained with ab32118 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab32118) (1x 106 in 100μl at 0.008μg/ml (1/258750)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
This antibody gave a positive signal in MCF7 Fixed with 4% formaldehyde (10 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.
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All lanes : Anti-Cytokeratin 18 antibody [E431-1] (ab32118) at 1/2000 dilution (Purified)
Lane 1 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 48 kDa
Observed band size: 48 kDa -
Immunocytochemistry/Immunofluorescence analysis of HeLa (+ve) and U87-MG (-ve) cells labelling Cytokeratin 18 with ab32118 at 2 ug/ml overnight at +4°C. Cells were fixed with 100% Methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. A preadsorbed Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150081) at 1/1000 dilution was used as the secondary antibody. The cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000), using ab150119, a preadsorbed Alexa Fluor® 647-conjugated goat anti-mouse IgG (1/1000) as the secondary antibody. Nuclei were counterstained with DAPI (blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 min).
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Immunocytochemistry/Immunofluorescence analysis of HT-29 cells labelling Cytokeratin 18 with purified ab32118 at 1/500. Cells were fixed with 100% Methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. The cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000) using ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary antibody. Nuclei were counterstained with DAPI (blue).
For negative control 1, rabbit primary antibody was used followed by anti-mouse secondary antibody (ab150120).
For negative control 2, mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) were used.Alexa Fluor® 488 (ab194124) conjugated version is available for this clone.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human stomach tissue sections labeling Cytokeratin 18 with purified ab32118 at 1/500 dilution (0.08 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Intracellular Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cytokeratin 18 with purified ab32118 at 1/20 dilution (2µg/ml) (red). Cells were fixed with 80% Methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluorr® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
R-PE (ab210410) conjugated version is available for this clone.
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Indirect immunofluorescence staining with antibodies against DNA, cytokeratin-7 and cytokeratin-18 (upper panel) or against DNA, vimentin and cytokeratin-18 (lower panel). Representatives are presented. Scale bar: 20 µm.
Control or treated cells were fixed for 15 min with 4% PFA containing 0.1% Triton X-100 at room temperature. The following primary antibodies were used for staining: monoclonal mouse antibodies against vimentin and cytokeratin-7 (both 1:100, DAKO) and monoclonal rabbit antibody against cytokeratin-18 (1:50, Abcam). DNA was stained using DAPI (4’,6-diamidino-2-phenylindole-dihydrochlorid) (Roche). Slides were examined using an Axio Imager 7.1 microscope (Zeiss) and images were taken using an Axio Cam MRm camera (Zeiss). The immunofluorescence stained slides were also examined by a confocal laser scanning microscope (CLSM) (Leica CTR 6500, Heidelberg). Images were processed using Photoshop.
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ab32118 showing positive staining in Normal colon tissue.
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Anti-Cytokeratin 18 antibody [E431-1] (ab32118) at 1/10000 dilution + A431 cell lysate
Predicted band size: 48 kDa
Observed band size: 48 kDa -
Overlay histogram showing MCF7 cells stained with ab32118 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32118, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF7 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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ab32118 showing positive staining in Gastric adenocarcinoma tissue.
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Fluorescent immunohistochemical analysis of paraffin-embedded human normal kidney tissue using ab32118. Green-CK18 red-PI
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ab32118 showing positive staining in Normal kidney tissue.
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ab32118 showing positive staining in Breast carcinoma tissue.
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ab32118 showing negative staining in Glioma tissue.
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ab32118 showing negative staining in Normal brain tissue.
数据表及文件
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SDS download
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Datasheet download
文献 (32)
ab32118 被引用在 32 文献中.
- Wang J et al. Exosomal-miR-10a derived from colorectal cancer cells suppresses migration of human lung fibroblasts, and expression of IL-6, IL-8 and IL-1ß. Mol Med Rep 23:N/A (2021). PubMed: 33236127
- Zheng X et al. microRNA-10a-5p overexpression suppresses malignancy of colon cancer by regulating human liver cancer fibroblasts. Neoplasma 68:1157-1168 (2021). PubMed: 34533029
- Tsering T et al. Uveal Melanoma-Derived Extracellular Vesicles Display Transforming Potential and Carry Protein Cargo Involved in Metastatic Niche Preparation. Cancers (Basel) 12:N/A (2020). PubMed: 33050649
- Chen YC et al. Mesenchymal Stem/Stromal Cell Engulfment Reveals Metastatic Advantage in Breast Cancer. Cell Rep 27:3916-3926.e5 (2019). PubMed: 31242423
- Djomehri SI et al. A reproducible scaffold-free 3D organoid model to study neoplastic progression in breast cancer. J Cell Commun Signal 13:129-143 (2019). PubMed: 30515709