Overview

  • Product nameAnti-Cytokeratin 18 antibody
    See all Cytokeratin 18 primary antibodies
  • Description
    Rabbit polyclonal to Cytokeratin 18
  • Tested applicationsSuitable for: ICC/IF, Flow Cyt, IP, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Cytokeratin 18 conjugated to Keyhole Limpet Haemocyanin (KLH) (Cysteine residue).
    (Peptide available as ab26325, ab26326)

  • Positive control
    • Hela, Jurkat and A431 cell lysates

Properties

Applications

Our Abpromise guarantee covers the use of ab24561 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
ICC/IF Use a concentration of 1 µg/ml.
Flow Cyt Use 1µg for 106 cells. ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
IP Use a concentration of 5 µg/ml.
IHC-P 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 48 kDa (predicted molecular weight: 48 kDa).

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 images

  • All lanes : Anti-Cytokeratin 18 antibody (ab24561) at 1 µg/ml

    Lane 1 : HeLa whole cell lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : A431 whole cell lysate (ab7909)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 48 kDa
    Observed band size : 48 kDa

    See Abreview

  • ICC/IF image of ab24561 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab24561, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
  • ab24561 (1/2000) staining Cytokeratin 18 in paraffin-embedded Human liver tissue.  Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval.  The secondary antibody was goat anti rabbit/mouse conjugated to HRP.  For further experimental details please refer to abreview.

    See Abreview

  • Overlay histogram showing MCF7 cells stained with ab24561 (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 (ab24561, 1µg/1x106 cells) 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 (polyclonal) (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.

  • Cytokeratin 18 was immunoprecipitated using 0.5mg A431 whole cell extract, 5µg of Rabbit polyclonal 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, A431 whole cell extract 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 ab24561.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 48kDa; Cytokeratin 18

References for Anti-Cytokeratin 18 antibody (ab24561)

This product has been referenced in:
  • Singh R  et al. Functional Analysis of Serially Expanded Human iPS Cell-Derived RPE Cultures. Invest Ophthalmol Vis Sci 54:6767-78 (2013). Human . Read more (PubMed: 24030465) »

See 1 Publication for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Liver)
Specification Liver
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Dako Target Retrieval Solution
Permeabilization No
Blocking step 5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C
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Verified customer

Submitted Mar 25 2010

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