Nori Human Cytokeratin 18 ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of cytokeratin 18 in human. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for CK18 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CK18 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for CK18 is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of CK18 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for cytokeratin 18: CK18, Keratin 18, KRT18
This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Human Cytokeratin 18 ELISA Kit Summary
Alternative names for cytokeratin 18: CK18, Keratin 18, KRT18
| Assay Type | Solid Phase Sandwich ELISA |
| Format | 96-well Microplate or 96-Well Strip Microplate |
| Method of Detection | Colorimetric |
| Number of Targets Detected | 1 |
| Target Antigen Accession Number |
P05783 |
| Assay Length | 3 hours |
| Quantitative/Semiquantitative | Quantitative |
| Sample Type | Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL, |
| Recommended Sample Dilution (Plasma/Serum) | No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ |
| Sensitivity | 6 pg/mL |
| Detection Range | 31.25-2000 pg/mL |
| Specificity | Human CK18 |
| Cross-Reactivity | < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested. |
| Interference | No significant interference observed with available related molecules |
| Storage/Stability | 4 ºC for up to 6 months |
| Usage | For Laboratory Research Use Only. Not for diagnostic or therapeutic use. |
| Additional Notes | The kit allows for use in multiple experiments. |
Standard Curve
Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer
Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water
Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.
Background:
Keratin 18 (KRT18) or cytokeratin 18 (CK18) is a type I cytokeratin and is encoded by KRT18 gene.[1] It is, together with its filament partner keratin 8, perhaps the most commonly found products of the intermediate filament gene family. They are expressed in single layer epithelial tissues of the body. Mutations in this gene have been linked to cryptogenic cirrhosis. Two transcript variants encoding the same protein have been found for this gene. the expression of CK18 was significantly downregulated in breast cancer tissues and in multiple drug resistance cell line overexpressing breast cancer resistance protein BCRP, and the presence of low levels of CK18 was associated with TNM stage, lymph node metastasis, and unfavorable survival. CK18 knockdown increased BCRP expression and induced the EMT process in human breast cancer MCF7 cells.[2] Serum CK18 is related strongly to the development/progression of NAFLD and HCV-related fibrosis/steatosis.[3] Subacute elevation of plasma caspase-cleaved cytokeratin-18 levels is associated with unfavorable early and late stroke outcome.[4] High KRT18 expression is associated with biliary tract cancer.[5] CK-18 serum levels were lower in longstanding type 1 diabetes patients compared to healthy controls.[6]Keratin 18 is often used together with keratin 8 and keratin 19 to differentiate cells of epithelial origin from hematopoietic cells in tests that enumerate circulating tumor cells in blood.[7]
References
- Romano,V., et al. (1986) Differentiation 30 (3), 244-253.
- Shi R, et al. (219) Oncol. Rep. 41 (5), 3015-3026.
- Darweesh SK, et al. (2019) Eur J Gastroenterol Hepatol 31 (5), 633-641.
- Molnar T, et al. (2019) J Stroke Cerebrovasc Dis 28 (3), 719-727.
- Sugimoto M, et al. (2018) Sci Rep 8 (1), 16997.
- Nurten E, et al. (2018) J. Pediatr. Endocrinol. Metab. 31 (9), 959-969.
- Jeffrey Allard; et al. (2004). Clin. Cancer Research. 10(20): 6897–6904.
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