Anti-Cytokeratin 18 antibody [E431-1] (ab32118)

Overview

Properties

Applications

Our Abpromise guarantee covers the use of ab32118 in the following tested applications.

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

Application Abreviews Notes
WB 1/10000. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).
IHC-P Use at an assay dependent concentration.
ICC/IF 1/100 - 1/500.

 

Flow Cyt 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent dilution.
  • Application notesIs unsuitable for IP.
  • Target

    • FunctionInvolved 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.
    • Tissue specificityExpressed in colon, placenta, liver and very weakly in exocervix. Increased expression observed in lymph nodes of breast carcinoma.
    • Involvement in diseaseDefects in KRT18 are a cause of cirrhosis (CIRRH) [MIM:215600].
    • Sequence similaritiesBelongs to the intermediate filament family.
    • Post-translational
      modifications
      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.
    • Cellular localizationCytoplasm > perinuclear region.
    • Information by UniProt
    • Database links
    • Alternative names
      • 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 [E431-1] images

    • Anti-Cytokeratin 18 antibody [E431-1] (ab32118) at 1/10000 dilution + A431 cell lysate

      Predicted band size : 48 kDa
      Observed band size : 48 kDa
    • 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.

    • ab32118 showing positive staining in Gastric adenocarcinoma tissue.

    • 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 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. 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.

    • Fluorescent immunohistochemical analysis of paraffin-embedded human normal kidney tissue using ab32118. Green-CK18 red-PI

    • ab32118 showing positive staining in Normal colon tissue.

    • ab32118 showing positive staining in Normal kidney tissue.

    • ab32118 showing positive staining in Breast carcinoma tissue.

    • ab32118 showing negative staining in Glioma tissue.

    • ab32118 showing negative staining in Normal brain tissue.

    References for Anti-Cytokeratin 18 antibody [E431-1] (ab32118)

    This product has been referenced in:
    • Lamichhane SP  et al. Recapitulating epithelial tumor microenvironment in vitro using three dimensional tri-culture of human epithelial, endothelial, and mesenchymal cells. BMC Cancer 16:581 (2016). IHC . Read more (PubMed: 27484993) »
    • Li J  et al. Reciprocal interaction of Wnt and RXR-a pathways in hepatocyte development and hepatocellular carcinoma. PLoS One 10:e0118480 (2015). Read more (PubMed: 25738607) »

    See all 14 Publications for this product

    Product Wall

    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (HeLa)
    Permeabilization Yes - 0.5% TritonX100 in PBS
    Specification HeLa
    Fixative Paraformaldehyde
    Username

    Dr. Kirk McManus

    Verified customer

    Submitted Jun 03 2015


    CK18
    ab32118
    86% homology - could detect the target signal at 1:1000 in mouse kidney and spleen using 5-fold more sensitive ECL substrate than the one we usually use.

    CK8
    ab32357
    89% homology - blank WB in mouse brain, hea...

    Read More

    Although mouse samples were not recommended based on the western blot results below with these antibodies, there is still the possibility that the negative results in mouse might be due to the limited sample types, especially for the antibodies with hi...

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    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Frozen sections)
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Sample Human Tissue sections (A549)
    Specification A549
    Permeabilization Yes - 0.5 Triton X-100
    Fixative Paraformaldehyde
    Username

    Mrs. Aleksandra Kotlyarova

    Verified customer

    Submitted Sep 26 2013

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Mouse Tissue sections (Pancreas)
    Specification Pancreas
    Fixative Paraformaldehyde
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrae pH 6
    Permeabilization No
    Blocking step BSA / FBS as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
    Username

    Mr. Raul Peña

    Verified customer

    Submitted Dec 03 2012

    Thank you very much for your answer.
    I have contacted the laboratory to gather some more information in regards to the species testing of these antibodies. I am sorry it has taken so long time. Have you already proceeded meanwhile?
    In regards...

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    Thank you for contacting us.
    I am sorry to hear this customer has been experiencing problems with 2 of our antibodies. Would it be possible to obtain more protocol information too so that we can investigate this in more detail?
    I look forward...

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    Thank you for your reply regarding ab32118.

    I am sorry that this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of ab668.

    To c...

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    Thank you for calling Abcam yesterday. I am sorry about the problems with ab32118. I have processed your request to receive ab668 as a free of charge replacement. Your new order number is ******. Please let me know if there is anything else I can help ...

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    Thank you for contacting Abcam. The concentration of ab32118 is approximately 0.130mg/ml and the concentration for ab32118 is 0.185 mg/ml. If there is anything else I can do, please do hesitate to ask.

    1-10 of 13 Abreviews or Q&A

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