FITC Anti-Cytokeratin 18 antibody [DC-10] (ab72813)

Submit a review Q&A (4)References (3)
Flow Cytometry - FITC Anti-Cytokeratin 18 antibody [DC-10] (ab72813)
  • Immunocytochemistry/ Immunofluorescence - FITC Anti-Cytokeratin 18 antibody [DC-10] (ab72813)

Key features and details

  • FITC Mouse monoclonal [DC-10] to Cytokeratin 18
  • Suitable for: Flow Cyt, ICC/IF
  • Reacts with: Human
  • Conjugation: FITC. Ex: 493nm, Em: 528nm
  • Isotype: IgG1

Overview

  • Product name

    FITC Anti-Cytokeratin 18 antibody [DC-10]
    See all Cytokeratin 18 primary antibodies

  • Description

    FITC Mouse monoclonal [DC-10] to Cytokeratin 18

  • Host species

    Mouse

  • Conjugation

    FITC. Ex: 493nm, Em: 528nm

  • Tested applications

    Suitable for: Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Human breast carcinoma cell line PMC-42.

  • Positive control

    • ICC/IF: HCT116 cells.

  • General notes

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab72813 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
Flow Cyt Use a concentration of 0.5 - 1 µg/ml.

ab91356 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF Use a concentration of 1 µg/ml.

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

    • Flow Cytometry - FITC Anti-Cytokeratin 18 antibody [DC-10] (ab72813)
      Flow Cytometry - FITC Anti-Cytokeratin 18 antibody [DC-10] (ab72813)
      Overlay histogram showing MCF-7 cells stained with ab72813 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab72813, 0.5µg/1x106 cells) for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 (FITC) (2µg/1x106 cells) for 30 min at 22°C. Acquisition of >5,000 events was performed.

    • Immunocytochemistry/ Immunofluorescence - FITC Anti-Cytokeratin 18 antibody [DC-10] (ab72813)
      Immunocytochemistry/ Immunofluorescence - FITC Anti-Cytokeratin 18 antibody [DC-10] (ab72813)
      ICC/IF image of ab72813 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 (ab72813, 1µg/ml) overnight at +4°C. 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.

    References (3)

    Publishing research using ab72813? Please let us know so that we can cite the reference in this datasheet.

    ab72813 has been referenced in 3 publications.

    • Zhang Q  et al. Isolation of circulating tumor cells and detection of EGFR mutations in patients with non-small-cell lung cancer. Oncol Lett 17:3799-3807 (2019). PubMed: 30881500
    • Zhang H  et al. A liquid biopsy-based method for the detection and quantification of circulating tumor cells in surgical osteosarcoma patients. Int J Oncol N/A:N/A (2017). PubMed: 28350107
    • Wagner BJ  et al. Phagocytosis of dying tumor cells by human peritoneal mesothelial cells. J Cell Sci 124:1644-54 (2011). PubMed: 21525033

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-4 of 4 Abreviews or Q&A

    Question

    We ordered the original antibody on 5/4/2012 and I have confirmation number ### I'd like to replace the antibody with the following polyclonal for cytokeratin 18: catalog # ab52948. Looking forward to trying the new antibody and thanks for your time.

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued an alternative free of charge replacement of ab52948:

    Order number: #####


    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    Question

    I checked the ab on our EVOS microscope, and what I loaded onto the slide is very fluorescent in the FITC channel, so I don't think the fluorochrome has dissociated from the antibody. Do you have any polyclonals that are FITC-conjugated? I'm wondering if this is too specific.

    Thanks,

    Read More
    Answer

    Thank you for taking the time to check the antibody under the microscope and get back to us. Your enquiry has now been forwarded to me as my colleague #### is currently away from the office.

    The details you have kindly provided will provide us with vital information for our monitoring of product quality.

    I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably received a bad vial. In this case, I am pleased to offer you a free of charge replacement or credit note in compensation if you have purchased within the 6 month guarantee period. In order to arrange this, I would appreciate if you are able to confirm the order number and date of purchase?

    If you prefer an alternative replacement, we do have many polyclonal cytokeratin 18 antibodies available. I have placed a link to the search results below. I can suggest to review the online datasheets for further information and to select one tested and therefore guaranteed in all the applications you are using.

    https://www.abcam.com/index.html?pageconfig=searchresults&search=cytokeratin 18&pt=1&sk=react&sv=21&sn=Human&l=5&fViewMore=1


    Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Question

    IHC Questionnaire It will take approximately 5 minutes to complete the questionnaire.
    Please fill-in all fields so that we can better assist you
    1) Abcam product code ab72813
    2) Abcam order reference number or product batch number 3) Description of the problem No positive staining on positive control cells
    4) Sample preparation:
    SpeciesHuman
    Type of sample: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other: cells in culture
    Sample preparationCells fixed on chamber slide
    Positive controlA431 cells (tested here)
    Negative controlNone
    5) Fixation step
    Yes/No Yes
    If yes: Fixative agent and concentration100% Methanol
    Fixation time 15 min
    Fixation temperature-20
    6) Antigen retrieval methodN/a
    7) Permeabilization method:methanol and Triton-X
    Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers? Yes
    Permeabilizing agent and concentration:
    100% methanol during fixation + 0.3% Triton-X blocking buffer and antibody dilution buffer
    8) Blocking agent (eg BSA, serum…):BSA
    Concentration 5%
    Blocking time 1 hour
    Blocking temperature RT
    9) Endogenous peroxidases blocked? NoEndogenous biotins blocked? No
    10) Primary antibody (If more than one was used, describe in “additional notes”) : n/aConcentration or dilution
    Diluent buffer
    Incubation time
    11) Secondary antibody:n/aSpecies:
    Reacts against:
    Concentration or dilution
    Diluent buffer
    Incubation time
    Fluorochrome or enzyme conjugate
    12) Washing after primary and secondary antibodies: Yes
    Buffer PBS
    Number of washes 3 x 5 min each
    13) Detection method EVOS microscope
    14) How many times have you run this staining?2x with A431 cells; 1x with mesenchymal stem cells
    Do you obtain the same results every time?Yes
    What steps have you altered to try and optimize the use of this antibody?Tried fixation with 10% formalin, followed by permeabilization with 0.1% Triton-X, andblocking buffer at 2% BSA
    Document attachment: Attaching images of your IHC is strongly recommended and can greatly speed up our investigation of your problem.
    Nopositivestaining

    Read More
    Answer

    Thank you very much for providing this information.

    From looking though the protocol used with Anti-Cytokeratin 18 antibody [DC-10] (FITC) (ab72813) I can see no reason why there would not be any signal observed in your cells.

    As discussed over the phone, I am seeking further information from the source of this antibody to see exactly how we have used this antibody in ICC.

    Could I just ask, have you tested that the FITC is still in tact? You could do this by loading on some of the antibody neat onto a slide and taking an image. Could I also ask how the antibody has been stored?

    I look forward to receiving your reply.

    Read More

    Question

    Phone calls reporting problems in using the Anti-Cytokeratin 18 antibody [DC-10] (FITC) (ab72813) in immunocytochemistry.

    Read More
    Answer

    Thank you for calling us today andalerting us to the problem you are experiencing with the Anti-Cytokeratin 18 antibody [DC-10] (FITC) (ab72813). We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

    As we discussed on the phone, Ihave attachedour questionnaire so that we can gather further information regarding the samples tested and the protocol used. OnceI havereceive the completed questionnaire,I will look at the protocol and see if there are any suggestionsI can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary.

    I am also currently looking for further information as to how the antibody has been used in immunocytochemistry previously and will share thisinformationwith you as soon as I have it.

    I look forward to receiving your reply.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com