Human Cytokeratin 18 Fragment ELISA Kit (ab254515)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 22.8 pg/ml
- Range: 46.875 pg/ml - 3000 pg/ml
- Sample type: Cell culture supernatant, Cell Lysate, Cit plasma, EDTA Plasma, Hep Plasma, Serum
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human Cytokeratin 18 Fragment ELISA Kit
See all Cytokeratin 18 kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Supernatant 8 2.2% Inter-assay Sample n Mean SD CV% Supernatant 3 9.5% -
Sample type
Cell culture supernatant, Serum, Cell Lysate, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
22.8 pg/ml -
Range
46.875 pg/ml - 3000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture supernatant 97 % - % Serum 87 % - % Cell Lysate 87 % - % Hep Plasma 79 % - % EDTA Plasma 85 % - % Cit plasma 84 % - % -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human Cytokeratin 18 Fragment ELISA Kit (ab254515) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Cytokeratin 18 Fragment protein in cell culture supernatant, cell lysate, cit plasma, edta plasma, hep plasma, and serum. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Cytokeratin 18 Fragment with 22.8 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
Cytokeratin 18 Fragment is a degradation product of Cytokeratin 18. Cytokeratin 18 is involved in the uptake of thrombin-antithrombin complexes by hepatic cells. Cytokeratin 18 becomes cleaved during apoptosis resulting in a 18kDa fragment, which can be used as a measure of cell death and a biomarker for liver disease. Ab245515 recognizes this fragment, and specifically recognizes the cleavage site (M30) neo-epitope and does not recognize the full-length protein.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human Cytokeratin 18 Fragment Capture Antibody 1 x 600µl 10X Human Cytokeratin 18 Fragment Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 4BI 1 x 6ml Human Cytokeratin 18 Fragment Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 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
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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
modificationsPhosphorylation 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
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Alternative names
- Cell proliferation inducing gene 46 protein
- Cell proliferation inducing protein 46
- Cell proliferation-inducing gene 46 protein
see all -
Database links
- Entrez Gene: 3875 Human
- Omim: 148070 Human
- SwissProt: P05783 Human
- Unigene: 406013 Human
Images
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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.
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The Cytokeratin 18 Fragment standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Cytokeratin 18 Fragment in human HeLa cell culture supernatant samples.
The concentrations of Cytokeratin 18 Fragment were measured in duplicates, interpolated from the Cytokeratin 18 Fragment standard curves and corrected for sample dilution. Undiluted samples are as follows: untreated 100% and STS treated 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Cytokeratin 18 Fragment concentration was determined to be 239 pg/mL in untreated and 400 pg/mL in STS treated cell culture supernatant.
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nterpolated concentrations of native Cytokeratin 18 Fragment in human HeLa cell extract and extract based on a 12.5 µg/mL and 1.56 µg/mL extract load (respectively).
The concentrations of Cytokeratin 18 Fragment were measured in duplicate and interpolated from the Cytokeratin 18 Fragment 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 Fragment concentration was determined to be 42.1 pg/mL / µg/mL in untreated and 394 pg/mL / µg/mL in STS treated HeLa cell extract.
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The concentrations of Cytokeratin 18 Fragment were measured in duplicates, interpolated from the Cytokeratin 18 Fragment standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 25%, plasma – citrate 25%, plasma- EDTA 25%, and plasma – heparin 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
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To learn more about the advantages of recombinant antibodies see here.