製品の概要

  • 製品名
    Anti-Cytokeratin 18 antibody [CK-18]
    Cytokeratin 18 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [CK-18] to Cytokeratin 18
  • 由来種
    Mouse
  • 特異性
    No cross reactivity with other proteins.
  • アプリケーション
    適用あり: Flow Cyt, WB, IHC-P, ICC/IF, IHC-Fr, IPmore details
  • 種交差性
    交差種: Rat, Human
  • 免疫原

    Tissue, cells or virus corresponding to Human Cytokeratin 18. Human epidermal carcinoma A431 and MCF7 human breast cancer cell lines.

  • ポジティブ・コントロール
    • This antibody gave a positive signal in HCT116 cell line in IF/ICC.
  • 特記事項

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab133263 as a replacement.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab82254 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

WB Use a concentration of 0.5 - 1 µg/ml. Predicted molecular weight: 48 kDa.
IHC-P Use a concentration of 1 - 2 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.
IHC-Fr Use a concentration of 0.5 - 2 µg/ml. Use formalin or acetone fixed tissues.
IP Use a concentration of 5 µg/ml.

ターゲット情報

  • 機能
    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.
  • 組織特異性
    Expressed in colon, placenta, liver and very weakly in exocervix. Increased expression observed in lymph nodes of breast carcinoma.
  • 関連疾患
    Defects in KRT18 are a cause of cirrhosis (CIRRH) [MIM:215600].
  • 配列類似性
    Belongs to the intermediate filament family.
  • 翻訳後修飾
    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.
  • 細胞内局在
    Cytoplasm > perinuclear region.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Cell proliferation inducing gene 46 protein antibody
    • Cell proliferation inducing protein 46 antibody
    • Cell proliferation-inducing gene 46 protein antibody
    • CK 18 antibody
    • CK-18 antibody
    • CK18 antibody
    • CYK 18 antibody
    • CYK18 antibody
    • Cytokeratin 18 antibody
    • Cytokeratin endo B antibody
    • Cytokeratin-18 antibody
    • K 18 antibody
    • K18 antibody
    • K1C18_HUMAN antibody
    • KA18 antibody
    • Keratin 18 antibody
    • Keratin 18, type I antibody
    • Keratin D antibody
    • keratin, type I cytoskeletal 18 antibody
    • Keratin-18 antibody
    • Krt18 antibody
    see all

画像

  • Anti-Cytokeratin 18 antibody [CK-18] (ab82254) + MCF-7 whole cell lysate

    Predicted band size: 48 kDa

  • ab82254 staining Cytokeratin 18 in paraffin-embedded human rectal cancer tissue.
  • ICC/IF image of ab82254 stained HCT116 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab82254, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • Overlay histogram showing MCF7 cells stained with ab82254 (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 (ab82254, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/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.

  • Cytokeratin 18 was immunoprecipitated using 0.5mg MCF7 Whole cell lysate, 5µg of Mouse monoclonal to Cytokeratin 18 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, MCF7 Whole cell lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab82254.
    Secondary: Protein G-HRP at 1/500 dilution.
    Band: 45kDa; Cytokeratin 18: non specific bands - 49kDa: We are unsure as to the identity of this extra band.

参考文献

This product has been referenced in:
  • Liu S  et al. Effect of Hypoxia on the Differentiation and the Self-Renewal of Metanephrogenic Mesenchymal Stem Cells. Stem Cells Int 2017:7168687 (2017). Flow Cyt . Read more (PubMed: 28194187) »
  • Yanagi Y  et al. In vivo and ex vivo methods of growing a liver bud through tissue connection. Sci Rep 7:14085 (2017). IHC ; Rat . Read more (PubMed: 29074999) »

See all 11 Publications for this product

レビューと Q&A

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab82254. I would also appreciate if you can confirm some further details: Before the protein blocking step we suggest use of a permeablization step. ...

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Thank you for your response.

It is important to mention that only ab133263 has been tested for IHC on formalin-fixed tissue samples. The other product (ab82254)is suitable for frozen sections, not been tested for formalin-fixed tissue samples...

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Thank you for your enquiry and your interest in our products.

Here are a few useful and interesting websites, you may wish to read through before starting the experiments:
http://docs.abcam.com/pdf/immunology/fluorochromes_table_updated....

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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