Overview

  • Product name

    Human Cytokeratin 18 ELISA Kit
    See all Cytokeratin 18 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 5 2.1%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 8.4%
  • Sample type

    Cell culture supernatant, Serum, Cell culture extracts, Tissue Extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    3.4 pg/ml
  • Range

    15.6 pg/ml - 1000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 100 99% - 102%
    Cell culture extracts 110 106% - 115%
    Cell culture media 109 107% - 110%
    Heparin Plasma 99 97% - 99%
    EDTA Plasma 99 97% - 100%
    Citrate Plasma 95 93% - 97%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat, Cow
  • Product overview

    Cytokeratin 18 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Cytokeratin 18 protein in human serum, plasma, cell culture supernatant, cell and tissue extract.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Cytokeratin is an intermediate filament and a marker of apoptosis. Cytokeratin 18 is also involved in the uptake of thrombin-antithrombin complexes by hepatic cells. Cytokeratin 18 can be cleaved into caspase cleaved Cytokeratin 18 fragments, which can be a measure of cell death and a biomarker for liver disease.


    Sensitivity:


    Samples diluted in Sample Diluent NS: 4.3 pg/mL.


    Samples diluted in 1X Cell Extraction Buffer PTR: 3.4 pg/mL.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 4BI 1 x 6ml
    10X Human Cytokeratin 18 Capture Antibody 1 x 600µl
    10X Human Cytokeratin 18 Detector Antibody 1 x 600µl
    Human Cytokeratin 18 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent LS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • 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
  • Alternative names

    • Cell proliferation inducing gene 46 protein
    • Cell proliferation inducing protein 46
    • Cell proliferation-inducing gene 46 protein
    • CK 18
    • CK-18
    • CK18
    • CYK 18
    • CYK18
    • Cytokeratin 18
    • Cytokeratin endo B
    • Cytokeratin-18
    • K 18
    • K18
    • K1C18_HUMAN
    • KA18
    • Keratin 18
    • Keratin 18, type I
    • Keratin D
    • keratin, type I cytoskeletal 18
    • Keratin-18
    • Krt18
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab227896 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Cytokeratin 18 were measured in duplicates, interpolated from the Cytokeratin 18 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (EDTA) 50%, plasma (heparin) 50% and plasma (citrate) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).   The mean Cytokeratin 18 concentration was determined to be 23 pg/mL in serum, 29pg/mL in plasma (EDTA), 96 pg/mL in plasma (heparin), and 33 pg/mL in plasma (citrate).

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). 5/10 donors were below the detectable dose. The mean of the detectable donors Cytokeratin 18 concentration was determined to be 59 pg/mL with a range of 31– 79 pg/mL.

  • The concentrations of Cytokeratin 18 were measured in duplicates, interpolated from the Cytokeratin 18 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (EDTA) 50%, plasma (heparin) 50% and plasma (citrate) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

  • The concentrations of Cytokeratin 18 were measured in duplicates, interpolated from the Cytokeratin 18 standard curves and corrected for sample dilution. Undiluted samples are as follows: Hela cell culture supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Cytokeratin 18 concentration was determined to be 1000 pg/mL in Hela cell culture supernatant.

  • The concentrations of Cytokeratin 18 were measured in duplicate and interpolated from the Cytokeratin 18 standard curve and corrected for sample dilution and extract load. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Cytokeratin 18 concentration was determined to be 0.89 pg / µg Hela extract and 35 pg/ µg Liver extract.

Protocols

References

ab227896 has not yet been referenced specifically in any publications.

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