Recombinant
RabMAb

Recombinant Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)

Reviews (2)Q&A (2)References (10)

Overview

  • Product name

    Anti-Cytokeratin 18 antibody [EPR1626]
    See all Cytokeratin 18 primary antibodies

  • Description

    Rabbit monoclonal [EPR1626] to Cytokeratin 18

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-Fr, Flow Cyt, WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Rat, Human

  • Immunogen

    Synthetic peptide corresponding to a region within the C-terminal end of Human Cytokeratin 18 (P05783).

  • Positive control

    • WB: HeLa, MCF7, and Rat liver, HeLa whole cell lysate (ab150035), Jurkat whole cell lysate (ab7899), and A431 cell lysates. IHC-P: human endometrium, Human gastric adenocarcinoma tissue, Human breast tissue, and rat liver tissues. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.

  • General notes

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab133263 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
IHC-Fr Use at an assay dependent dilution.
Flow Cyt 1/20.
WB 1/1000. Detects a band of approximately 45 kDa (predicted molecular weight: 48 kDa).

For unpurified use at 1/10,000 - 1/50,000
IHC-P 1/500 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Perform antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols.

1:500 in human tissues / 1:1000 in rat tissues
ICC/IF 1/50.

For unpurified use at 1/250 - 1/500
  • Application notes
    Is unsuitable for IP.

    • Target

    • Function

      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.

    • Tissue specificity

      Expressed in colon, placenta, liver and very weakly in exocervix. Increased expression observed in lymph nodes of breast carcinoma.

    • Involvement in disease

      Defects in KRT18 are a cause of cirrhosis (CIRRH) [MIM:215600].

    • Sequence similarities

      Belongs 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 localization

      Cytoplasm > 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

      Images

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human endometrium tissue sections labeling Cytokeratin 18 with Purified ab133263 at 1:500 dilution (0.23 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Purified)ImmunoHistoProbe one step HRP Polymer (ready to use) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
      • Flow Cytometry - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Flow Cytometry - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)

        Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Cytokeratin 18 with Purified ab133263 at 1:20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 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).

      • Western blot - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Western blot - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        All lanes : Anti-Cytokeratin 18 antibody [EPR1626] (ab133263) at 1/20000 dilution (Purified)

        Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
        Lane 2 : MCF7 (Human breast 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: 45 kDa
        why is the actual band size different from the predicted?

      • Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)

        Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Cytokeratin 18 with Purified ab133263 at 1:50 dilution (2.26 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

        Alexa Fluor® 488 (ab206091) and Alexa Fluor® 647 (ab206269) conjugated versions are available for this clone.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling Cytokeratin 18 with Purified ab133263 at 1:1000 dilution (0.113 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Purified)ImmunoHistoProbe one step HRP Polymer (ready to use) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
      • Western blot - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Western blot - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Anti-Cytokeratin 18 antibody [EPR1626] (ab133263) at 1/1000 dilution (Purified) + Rat liver lysates at 20 µg

        Secondary
        Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution

        Predicted band size: 48 kDa
        Observed band size: 45 kDa why is the actual band size different from the predicted?

      • Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunofluorescence analysis of HeLa cells labelling Cytokeratin 18 with ab133263 at 1/250.
      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)

        Immunohistochemical analysis of paraffin embedded Human gastric adenocarcinoma tissue labelling Cytokeratin 18 with ab133263 at 1/250.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Western blot - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Western blot - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        All lanes : Anti-Cytokeratin 18 antibody [EPR1626] (ab133263) at 1/10000 dilution

        Lane 1 : MCF7 cell lysate
        Lane 2 : HeLa cell lysate
        Lane 3 : Jurkat cell lysate
        Lane 4 : A431 cell lysate

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

        Predicted band size: 48 kDa

      • Flow Cytometry - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Flow Cytometry - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)

        Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Cytokeratin 18 with unpurified ab133263 at 1/20 dilution(10ug/ml) (red). Cells were fixed with 80% methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor® 488)(ab150077)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black)(ab172730) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)

        Immunohistochemical analysis of paraffin embedded Human breast tissue labelling Cytokeratin 18 with ab133263 at 1/250.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)

        ab133263 showing positive staining in Prostatic carcinoma tissue.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)

        ab133263 showing negative staining in Skeletal muscle tissue.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)

        ab133263 showing negative staining in Normal brain tissue.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)

        ab133263 showing positive staining in Breast carcinoma tissue.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)

        ab133263 showing positive staining in Lung adenocarcinoma tissue.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 18 antibody [EPR1626] (ab133263)

        ab133263 showing negative staining in Cervical squamous carcinoma tissue.

        Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

      References

      This product has been referenced in:

      • Chen X  et al. Increase of glandular epithelial cell clusters by an external volume expansion device promotes adipose tissue regeneration by recruiting macrophages. Biosci Rep 39:N/A (2019). Read more (PubMed: 30760630) »
      • Zhang J  et al. KRT18 is correlated with the malignant status and acts as an oncogene in colorectal cancer. Biosci Rep 39:N/A (2019). Read more (PubMed: 31345960) »
      See all 10 Publications for this product

      Publishing research using ab133263? Please let us know so that we can cite the reference in this datasheet.

      Customer reviews and Q&As

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      1-4 of 4 Abreviews or Q&A

      Immunocytochemistry/ Immunofluorescence abreview for Anti-Cytokeratin 18 antibody [EPR1626]

       Excellent
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (HeLa)
      Permeabilization
      Yes - 0.5% TritonX100 in PBS
      Specification
      HeLa
      Fixative
      Paraformaldehyde
      Read More

      Dr. Kirk Mcmanus

      Verified customer

      Submitted Apr 22 2015

      Immunohistochemistry (Frozen sections) abreview for Anti-Cytokeratin 18 antibody [EPR1626]

       Good
      Abreviews
      abreview image
      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 cell line)
      Specification
      A549 cell line
      Permeabilization
      Yes - 0.5% Triton X100
      Fixative
      Paraformaldehyde
      Read More

      Mrs. Aleksandra Kotlyarova

      Verified customer

      Submitted Sep 26 2013

      Question

      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

      Read More
      Answer

      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.

      I hope this helps and if I can assist further, please do not hesitate to contact me.

      Read More

      Question

      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 [EPR1626] (ab133263)) could be right choice for my purpose? Could you please confirm to me if I am right or if there is better choice could you please recommend to me? I am microbiologist and my background in histology is not very good … that is why I need a complete product to use directly for the specific stain (the antibody with the fluorescent stain together) is it available such product with the condition I request above (fixed rat mammary tissue at Ex/Em 632/650)? My last aim is to get a red fluorescent photo for the tissue like the ones you present in the file of the two items ((ab133263) and (ab82254)) please send to me your answer as soon as possible? Many thanks in advance

      Read More
      Answer

      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:
      https://docs.abcam.com/pdf/immunology/fluorochromes_table_updated.pdf
      https://www.abcam.com/index.html?pageconfig=resource&rid=12922
      https://www.abcam.com/index.html?pageconfig=popular_protocols
      Both antibodies are suitable for detecting rat cytokeratin 18 and have been tested for immunohistochemistry. You may need to consider how you wish to fix the samples i.e. formalin, acetone, methanol, ethanol etc.









      These antibodies are unconjugated, so you need a compatible secondary antibody which is conjugated for the detection. In order to be able to select the right secondary antibody, it is essential to understand the isotype and the host species of your chosen primary antibody:

      - ab82254: mouse IgG1 so the secondary should be anti-mouse IgG1;

      - ab133263: rabbit IgG1 so the secondary should be anti-rabbit IgG1;

      I wouldrecommend conducting a search for the secondary antibody using the advance search engine. You can select the target species, the host species, isotype, format, conjugate etc:

      https://www.abcam.com/index.html?pageconfig=productmap&cl=918

      I hope this helps and if I can assist further, please do not hesitate to contact me.

      Read More

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      For licensing inquiries, please contact partnerships@abcam.com