Anti-Cytokeratin 18抗体[CK-18] (ab82254)

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概述

  • 产品名称
    Anti-Cytokeratin 18抗体[CK-18]
    参阅全部 Cytokeratin 18 一抗
  • 描述
    小鼠单克隆抗体[CK-18] to Cytokeratin 18
  • 特异性
    No cross reactivity with other proteins.
  • 经测试应用
    适用于: Flow Cyt, WB, IHC-P, ICC/IF, IHC-Fr, IPmore details
  • 种属反应性
    与反应: 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.

  • 阳性对照
    • This antibody gave a positive signal in HCT116 cell line in IF/ICC.

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 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
  • 数据库链接
    • 别名
      • 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

    Anti-Cytokeratin 18 antibody [CK-18] 图像

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

      Predicted band size : 48 kDa
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Cytokeratin 18 antibody [CK-18] (ab82254)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Cytokeratin 18 antibody [CK-18] (ab82254)
      ab82254 staining Cytokeratin 18 in paraffin-embedded human rectal cancer tissue.
    • Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 18 antibody [CK-18] (ab82254)
      Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 18 antibody [CK-18] (ab82254)
      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.
    • Flow Cytometry-Anti-Cytokeratin 18 antibody [CK-18](ab82254)
      Flow Cytometry-Anti-Cytokeratin 18 antibody [CK-18](ab82254)
      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.

    • Immunoprecipitation - Anti-Cytokeratin 18 antibody [CK-18] (ab82254)
      Immunoprecipitation - Anti-Cytokeratin 18 antibody [CK-18] (ab82254)
      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.

    Anti-Cytokeratin 18 antibody [CK-18] (ab82254)参考文献

    This product has been referenced in:
    • Powell E  et al. p53 deficiency linked to B cell translocation gene 2 (BTG2) loss enhances metastatic potential by promoting tumor growth in primary and metastatic sites in patient-derived xenograft (PDX) models of triple-negative breast cancer. Breast Cancer Res 18:13 (2016). IHC ; Human . Read more (PubMed: 26818199) »
    • Kruithof-de Julio M  et al. Canonical Wnt signaling regulates Nkx3.1 expression and luminal epithelial differentiation during prostate organogenesis. Dev Dyn 242:1160-71 (2013). Read more (PubMed: 23813564) »

    See all 2 Publications for this product

    发表研究结果有使用 ab82254？请让我们知道，以便我们可以引用本数据表中的参考文章。

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    3 Abreviews or Q&A

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    Votes: Highest First

    I am not seeing any staining in ICC/IF.

    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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    Thanks for your replay and all the useful information
    I have already fixed all the tissue with formalin 4% … is it possible to
    use the antibody for fixed tissue?
    Thanks

    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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    Inquiry: Dear Sir or Madam I want to stain a fixed rat mammary gland tissue with fluorescent dye have excitation and emission wave length between 632/650 I thought (Anti-Cytokeratin 18 antibody [CK-18] (ab82254)) or (Anti-Cytokeratin 18 antibody [EPR16...

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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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