categories: Cytoskeleton: Intermediate Filaments, Cytoskeleton: Intermediate Filaments (Human)

Mouse Monoclonal to Cytokeratin 18

DA-7 (IgG1)

Product No.	Form	Quantity	Price
11-110-C025	purified	0.025 mg	choose region
11-110-C100	purified	0.1 mg	choose region
1B-110-C025	biotin	0.025 mg	choose region
1B-110-C100	biotin	0.1 mg	choose region

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Background

Cytokeratins are a subfamily of intermediate filaments and characterized by remarkable biochemical diversity. Cytokeratins are represented in epithelial tissues by at least 20 different polypeptides, molecular weight between 40 kDa and 68 kDa. The individual cytokeratin polypeptides are designated 1 to 20 and divided into the type I (acidic cytokeratins 9-20) and type II (basic to neutral cytokeratins 1-8) families.
Cytokeratins 18 belongs to type I family (acidic cytokeratins).

Entrez Gene (cytokeratin 18, human): 12q13

Specificity

The antibody DA-7 reacts with Cytokeratin 18 (45 kDa). Cytokeratins are a member of intermediate filaments subfamily represented in epithelial tissues.

Immunogen

Human breast carcinoma cell line PMC-42.

Species Reactivity: Human Negative Species: Porcine Mouse Rat Hamster Bovine Canine (Dog) Sheep	Applications: Immunoprecipitation Western Blotting Recommended dilution: 1-2 μg/ml, overnight in 4°C Positive control: HeLa human cervix carcinoma cell line A-431 human epidermoid carcinoma cell line MCF-7 human breast adenocarcinoma cell line Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold Lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate with non-reducing SDS-PAGE sample buffer. Application note: Non-reducing conditions. SDS-PAGE (10% separating gel). Immunohistochemistry (paraffin sections) Recommended dilution: 10 μg/ml Positive tissue: colon, skin Immunocytochemistry Positive control: MCF-7 human breast adenocarcinoma cell line HeLa human cervix carcinoma cell line ELISA Positive control: MCF-7 human breast adenocarcinoma cell line Application note: The antibody DA-7 has been tested as the detection antibody in a sandwich ELISA for analysis of Cytokeratin 18 in combination with the antibody C-04 (cat. no. 11-106-C100). Usage note: Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.	General references Taylor-Papadimitriou J, Stampfer M, Bartek J, Lewis A, Boshell M, Lane EB, Leigh IM: Keratin expression in human mammary epithelial cells cultured from normal and malignant tissue: relation to in vivo phenotypes and influence of medium. J Cell Sci. 1989 Nov;94 ( Pt 3):403-13. [Abstract] Kovarik J. et al.: J. Tumor Marker Oncol. 5, 219 (1990). Product Specific References Lauerova L, Kovarik J, Bartek J, Rejthar A, Vojtesek B: Novel monoclonal antibodies defining epitope of human cytokeratin 18 molecule. Hybridoma. 1988 Oct;7(5):495-504. [Abstract] Kovarik J, Rejthar A, Lauerova L, Vojtesek B, Bartkova J: Monoclonal antibodies against individual cytokeratins in the detection of metastatic spread. Int J Cancer Suppl. 1988;3:50-5. [Abstract] *Vojtesek B, Staskova Z, Nenutil R, Lauerova L, Kovarik J, Rejthar A, Bartkova J, Bartek J: Monoclonal antibodies recognizing different epitopes of cytokeratin No.18. Folia Biol (Praha). 1989;35(6):373-82. [Abstract]

Species Reactivity:

Human

Negative Species:

Porcine
Mouse
Rat
Hamster
Bovine
Canine (Dog)
Sheep

Applications:

Immunoprecipitation
Western Blotting
Recommended dilution:
1-2 μg/ml, overnight in 4°C
Positive control:
HeLa human cervix carcinoma cell line
A-431 human epidermoid carcinoma cell line
MCF-7 human breast adenocarcinoma cell line
Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold Lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate with non-reducing SDS-PAGE sample buffer.
Application note: Non-reducing conditions. SDS-PAGE (10% separating gel).
Immunohistochemistry (paraffin sections)
Recommended dilution: 10 μg/ml
Positive tissue: colon, skin
Immunocytochemistry
Positive control:
MCF-7 human breast adenocarcinoma cell line
HeLa human cervix carcinoma cell line
ELISA
Positive control: MCF-7 human breast adenocarcinoma cell line
Application note: The antibody DA-7 has been tested as the detection antibody in a sandwich ELISA for analysis of Cytokeratin 18 in combination with the antibody C-04 (cat. no. 11-106-C100).

Usage note:
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.

General references

*Taylor-Papadimitriou J, Stampfer M, Bartek J, Lewis A, Boshell M, Lane EB, Leigh IM: Keratin expression in human mammary epithelial cells cultured from normal and malignant tissue: relation to in vivo phenotypes and influence of medium. J Cell Sci. 1989 Nov;94 ( Pt 3):403-13. [Abstract]

*Kovarik J. et al.: J. Tumor Marker Oncol. 5, 219 (1990).

Product Specific References

*Lauerova L, Kovarik J, Bartek J, Rejthar A, Vojtesek B: Novel monoclonal antibodies defining epitope of human cytokeratin 18 molecule. Hybridoma. 1988 Oct;7(5):495-504. [Abstract]

*Kovarik J, Rejthar A, Lauerova L, Vojtesek B, Bartkova J: Monoclonal antibodies against individual cytokeratins in the detection of metastatic spread. Int J Cancer Suppl. 1988;3:50-5. [Abstract]

*Vojtesek B, Staskova Z, Nenutil R, Lauerova L, Kovarik J, Rejthar A, Bartkova J, Bartek J: Monoclonal antibodies recognizing different epitopes of cytokeratin No.18. Folia Biol (Praha). 1989;35(6):373-82. [Abstract]

For research use only. Not for drug, diagnostic or other use.

Related Products

Mouse IgG1 Isotype Control

Mouse Monoclonal to Cytokeratin 18 C-04 (IgG1)

Mouse Monoclonal to Cytokeratin 18 DC-10 (IgG1)

Example Data

Fig. 1. Western Blotting

Fig. 1. Western Blotting analysis (reducing conditions) of whole cell lysate using anti-human Cytokeratin 18 (DA-7).
Lane 1: HeLa human cervix carcinoma cell line
Lane 2: A-431 human epidermoid carcinoma cell line
Lane 3: MCF-7 human breast adenocarcinoma cell line

Fig. 2.

Fig. 2. Immunohistochemistry staining of human colon (paraffin sections) using anti-cytokeratin 18 (clone DA-7).
Commercially tested by LifeSpan BioSciences.

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