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Human Cytokeratin 18 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF7619

R&D Systems, part of Bio-Techne
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AF7619
AF7619-SP

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunocytochemistry, Immunohistochemistry, Knockout Validated, Simple Western, Western Blot

Cited:

Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG

Product Specifications

Immunogen

E. coli-derived recombinant human Cytokeratin 18
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 antibody by Western Blot.

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 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

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.

Applications for Human Cytokeratin 18 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
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

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

Western Blot

0.1-1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and A431 human epithelial carcinoma cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.

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.

Reconstitution Buffer Available:
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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.

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.

Alternate Names

CK-18, CYK18, Keratin 18, KRT18

Entrez Gene IDs

3875 (Human); 16668 (Mouse)

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 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

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