Human Cytokeratin 18 Antibody

Catalog # Availability Size / Price Qty
AF7619
AF7619-SP
Detection of Human Cytokeratin 18 by Western Blot.
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Product Details
Citations (2)
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Human Cytokeratin 18 Antibody Summary

Species Reactivity
Human
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.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human Cytokeratin 18
Ala239-Asp397
Accession # P05783
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1-1 µg/mL
See below
Simple Western
1 µg/mL
See below
Immunohistochemistry
3-15 µg/mL
See below
Immunocytochemistry
5-15 µg/mL
See below
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.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Human Cytokeratin 18 antibody by Western Blot. View Larger

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.

Western Blot View Larger

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.

Immunocytochemistry Cytokeratin 18 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC). View Larger

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.

Immunohistochemistry Cytokeratin 18 antibody in Human Breast Cancer Tissue by Immunohistochemistry (IHC-P). View Larger

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.

Simple Western Detection of Human Cytokeratin 18 antibody by Simple Western<sup>TM</sup>. View Larger

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.

Knockout Validated View Larger

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.

Immunocytochemistry View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
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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.

Background: Cytokeratin 18

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.

Entrez Gene IDs
3875 (Human); 16668 (Mouse)
Alternate Names
Cell proliferation-inducing gene 46 protein; cell proliferation-inducing protein 46; CK-18; CYK18; Cytokeratin 18; cytokeratin-18; K18; Keratin 18; keratin, type I cytoskeletal 18; keratin-18; KRT18

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Citations for Human Cytokeratin 18 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. TFAP2C- and p63-Dependent Networks Sequentially Rearrange Chromatin Landscapes to Drive Human Epidermal Lineage Commitment
    Authors: L Li, Y Wang, JL Torkelson, G Shankar, JM Pattison, HH Zhen, F Fang, Z Duren, J Xin, S Gaddam, SP Melo, SN Piekos, J Li, EJ Liaw, L Chen, R Li, M Wernig, WH Wong, HY Chang, AE Oro
    Cell Stem Cell, 2019-01-24;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  2. Retinoic acid and BMP4 cooperate with p63 to alter chromatin dynamics during surface epithelial commitment
    Authors: JM Pattison, SP Melo, SN Piekos, JL Torkelson, E Bashkirova, MR Mumbach, C Rajasingh, HH Zhen, L Li, E Liaw, D Alber, AJ Rubin, G Shankar, X Bao, HY Chang, PA Khavari, AE Oro
    Nat. Genet., 2018-11-05;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC

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