Cytokeratin 18; also KRT-18 (Keratin, type I cytoskeletal 18), Cell proliferation-inducing gene 46 and Keratin-18) is a 44-46 kDa Class I (large keratins of acidic pH) member of the intermediate filament family of proteins. Individual keratins are always expressed in tandem with a second keratin, and these are found in all epithelial cells. The class I Cytokeratin 18 heterodimerizes/polymerizes with 50-52 kDa class II KRT-8 to form 8-10 nm filaments in single strata plus hepatic epithelia. Cytokeratin 18 and -8 are the first keratins to appear in the mammalian embyro. In the adult, Cytokeratin 18 appears to participate in subtractions and additions to the plasma membrane. In this regard, a number of intracellular proteins interact with Cytokeratin 18, including 14-3-3, HSPc70 and Mrj. Cytokeratin 18 may also be O‑glycosylated, and when so, serves to promote Akt-1 activity, thus protecting against apoptosis. Human Cytokeratin 18 is 430 amino acids (aa) in length. It contains an N-terminal "head" region (aa 1-79), a subsequent "rod" region (aa 80-387) with two coiled segments, and a C-terminal tail region. Cytokeratin 18 possesses at least 19 utilized phosphorylation sites plus five acetylated Lys residues. There are multiple isoforms that range from 20-40 kDa in size and are the result of caspase cleavage. A principal cleavage site occurs after Asp238. Over aa 239-397, human Cytokeratin 18 shares 86% aa sequence identity with mouse Cytokeratin 18.
Human Cytokeratin 18 Antibody
R&D Systems | Catalog # AF7619
Key Product Details
Validated by
Knockout/Knockdown
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
Knockout Validated, Immunohistochemistry, Western Blot, Immunocytochemistry, Simple Western
Cited:
Immunocytochemistry
Label
Unconjugated
Antibody Source
Polyclonal Sheep IgG
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Product Specifications
Immunogen
E. coli-derived recombinant human Cytokeratin 18
Ala239-Asp397
Accession # P05783
Ala239-Asp397
Accession # P05783
Specificity
Detects human Cytokeratin 18 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human Cytokeratin 14 (KRT14) is observed.
Clonality
Polyclonal
Host
Sheep
Isotype
IgG
Scientific Data Images for Human Cytokeratin 18 Antibody
Detection of Human Cytokeratin 18 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and A431 human epithelial carcinoma cell line. PVDF membrane was probed with 0.1 µg/mL of Sheep Anti-Human Cytokeratin 18 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7619) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for Cytokeratin 18 at approximately 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.Detection of Human Cytokeratin 18 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line (Cytokeratin 18 positive) and MOLT‑4 human acute lymphoblastic leukemia cell line (Cytokeratin 18 negative). PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Cytokeratin 18 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7619) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for Cytokeratin 18 at approximately 45 kDa (as indicated). GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.Cytokeratin 18 in HeLa Human Cell Line.
Cytokeratin 18 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human Cytokeratin 18 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7619) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoskeletal fibers. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.Cytokeratin 18 in Human Breast Cancer Tissue.
Cytokeratin 18 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Sheep Anti-Human Cytokeratin 18 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7619) at 3 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to intermediate filaments. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.Detection of Human Cytokeratin 18 by Simple WesternTM.
Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and A431 human epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Cytokeratin 18 at approximately 57 kDa (as indicated) using 1 µg/mL of Sheep Anti-Human Cytokeratin 18 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7619) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.Western Blot Shows Human Cytokeratin 18 Specificity by Using Knockout Cell Line.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and human KRT-18 knockout HeLa human cervical epithelial carcinoma cell line (KO). PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Cytokeratin 18 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7619) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for Cytokeratin 18 at approximately 48 kDa (as indicated) in the parental HeLa human cervical epithelial carcinoma cell line, but is not detectable in knockout HeLa human cervical epithelial carcinoma cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.Detection of Cytokeratin 18 in HeLa Cells (Positive) and HeLa KRT18 Knockout (Negative) Control.
Cytokeratin 18 was detected in immersion fixed HeLa Human Cervical Epithelial Carcinoma Cells (Positive) and absent in HeLa KRT18 Knockout (Negative) Control using Sheep Anti-Human Cytokeratin 18 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7619) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.Applications for Human Cytokeratin 18 Antibody
Application
Recommended Usage
Immunocytochemistry
5-15 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line
Immunohistochemistry
3-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human breast cancer tissue
Sample: Immersion fixed paraffin-embedded sections of human breast cancer tissue
Knockout Validated
Cytokeratin
18 is specifically detected in HeLa human cervical epithelial carcinoma
parental cell line but is not detectable in Cytokeratin 18 knockout HeLa
human cervical epithelial carcinoma cell line.
Simple Western
1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and A431 human epithelial carcinoma cell line
Sample: HeLa human cervical epithelial carcinoma cell line and A431 human epithelial carcinoma cell line
Western Blot
0.1-1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and A431 human epithelial carcinoma cell line
Sample: HeLa human cervical epithelial carcinoma cell line and A431 human epithelial carcinoma cell line
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: Cytokeratin 18
Alternate Names
CK-18, CYK18, Keratin 18, KRT18
Gene Symbol
KRT18
UniProt
Additional Cytokeratin 18 Products
Product Documents for Human Cytokeratin 18 Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human Cytokeratin 18 Antibody
For research use only
Related Research Areas
Citations for Human Cytokeratin 18 Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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