Antibody keratin

Double-label ICC is another approach using a similar principle. It pairs a nuclear antibody with a cytoplasmic antibody on the same sample, such as keratin and estrogen receptor. At present, this method is mostly used in the research field. 

Altmannsberger M, Dirk T, Droese M, Weber K, Osborn M (1986) Keratin polypeptide distribu tion in benign and malignant breast tumors subdivision of ductal carcinomas using monoclo nal antibodies. Virchows Arch B Cell Pathol Incl Mol Pathol 51(3) 265 275 Assefnia S, Jones LP, Torre KM, Furth PA (2006) Expression of p63 in a mouse model of Brcal mutation related breast cancer. Poster presented at the Annual Meeting of the American Association for Cancer Research, Washington, DC (Meeting Abstracts)... 

Bartek J, Vojtesek B, Staskova Z, Bartkova J, Kerekes Z, Rejthar A, Kovarik J (1991) A series of 14 new monoclonal antibodies to keratins characterization and value in diagnostic histopa thology. J Pathol 164(3) 215 224... 

Ivanyi D, Ansink A, Groeneveld E, Hageman PC, Mooi WJ, Heintz AP (1989) New monoclonal antibodies recognizing epidermal differentiation associated keratins in formalin fixed, paraffin embedded tissue. Keratin 10 expression in carcinoma of the vulva. J Pathol 159... 

Moll R, Franke WW, Vole Platzer B, Krepler R (1982b) Different keratin polypeptides in epider mis and other epithelia of human skin a specific cytokeratin of molecular weight 46,000 in epithelia of the pilosebaceous tract and basal cell epitheliomas. J Cell Biol 95(1) 285 295 Moll R, Krepler R, Franke WW (1983) Complex cytokeratin polypeptide patterns observed in certain human carcinomas. Differentiation 23(3) 256 269 Moll R, Dhouailly D, Sun TT (1989) Expression of keratin 5 as a distinctive feature of epithelial and biphasic mesotheliomas. An immunohistochemical study using monoclonal antibody AE14. Virchows Arch B Cell Pathol Incl Mol Pathol 58(2) 129 145 Moll R, Schiller DL, Franke WW (1990) Identification of protein IT of the intestinal cytoskeleton as a novel type I cytokeratin with unusual properties and expression patterns. J Cell Biol 111(2) 567 580... 

Moll R, Lowe A, Laufer J, Franke WW (1992) Cytokeratin 20 in human carcinomas. A new histodiagnostic marker detected by monoclonal antibodies. Am J Pathol 140 427 447 Moll R, Divo M, Langbein L (2008) The human keratins biology and pathology. Histochem Cell Biol 129 705 733... 

There are 20 amino acids found in the human body that are involved in protein synthesis. Refer to your textbook for a table of the 20 amino acids. The sequence of the amino acids dictates the properties of a protein. Examples of proteins include keratin in hair, hemoglobin, insulin, antibodies, and enzymes. 

Figure 26.5 Immunofluorescent staining for transformed airway epithelial cells with antibodies specific for (A) keratin 18 (a marker of epithelial cells) and (B) ZO-1 (marker of tight junction formation). Both markers indicate that the cells have retained epithelial characteristics after transformation. The staining for the presence of ZO-1 at the periphery of the cells indicates that the cells have not lost their polarity and can form tight monolayers that will generate a transepithelial resistance. This is a particularly attractive feature for the analysis of ion transport and transcellular transport of macromolecules. ZO-1 is also found in the nucleus and can also be detected by the anti-ZO-1 antibody.

Figure 6. Cytoskeletal proteins in nucleus. HeLa cells were immunostained with anti-keratin 8 or anti-keratin 18 antibodies, (a) Paraformaldehyde-fixed cells, (b) Cells treated with detergent before the fixation. (Scale bars 10pm)...

An abbreviated panel of antibodies directed against three antigens—keratin, S-100 protein, and CD45 (leukocyte common antigen—EC A)—can separate... 

This antihypertensive drug was withdrawn from use after a number of cases of occulomucocutaneous syndrome (skin rashes, keratinization of the eye, and other tissues). Mechanism is unknown, but there is evidence for metabolism (microsomal enzyme mediated) to a reactive intermediate capable of reacting with protein and forming antigenic conjugates. Antipractolol antibodies are detected in patients. Syndrome is not reproducible in animals, but in hamsters, practolol accumulates in the eye. 

Sondell, B., Thomell, L.-E., Stigbrand, T., and Egelrud, T., Immunolcoalization of stratum corneum chymotryptic enzyme in human skin and oral epithelium with monoclonal antibodies evidence of a proteinase specifically expressed in keratinizing squamous epithelia, J. Histochem. Cytochem., 42, 459, 1994. 

A direct detection method was recently developed for these adducts in stratum comeum of human skin based on immunofluorescence microscopy (30). Three partial sequences of keratins containing glutamine or asparagine, adducted with a 2-hydroxyethyl-thioethyl group at the omega-amide function, were synthesized and used as antigens for raising antibodies. After immunization, monoclonal antibodies were obtained with affinity for keratin isolated from human callus exposed to 50 xM sulfur mustard (see Plate 1). In contrast to the immunochemical... 

Abnormal tissues The antibody labeled 17/20 sarcomas, 16/18 melanomas, 4/4 meningeomas, and 3/3 schwannomas, and was the sole intermediate filament present in these tumours. In addition, variable percentages (10 to 57 percent) of carcinomas, neuroendocrine carcinomas, neuroblastomas, thymomas and mesotheliomas were positive with the antibody. With the exception of the neuroblastomas, cytokeratin was coexpressed with vimentin in these tumours. Among adenocarcinomas, more than 50 percent of papillary carcinomas of the thyroid as well as renal, endometrial, ovarian and lung carcinomas were labeled by the antibody and coexpressed keratins and vimentin. 

An abbreviated panel of antibodies directed against three antigens— keratin, S-100 protein, and CD45 (Leukocyte Common Antigen-LCA)— can separate poorly differentiated tumors into four diagnostic categories carcinoma, neural tissue tumor/melanoma, hematopoietic tumor, and sarcoma as shown in Table 2. 

Antibodies to these PEMs are directed to the apical membrane portions of secretory mammary cells surrounding milk fat globules (mol wt >250 kDa). They react with a variety of epithelial tumors, including mesotheliomas. Like keratin, these antibodies are also expressed on synovial sarcomas, epithelioid sarcomas, and chordomas. They are not as reactive with as wide a variety of tumors as antikeratins. For instance, they do not react with many endocrine tumors. However, they may recognize some renal-cell carcinomas and small-cell anaplastic carcinomas... 

Based on size and shape, the proteins are also subdivided into globular and fibrous proteins. Globular proteins are mostly water-soluble and fragile in nature. Example- enzymes, hormones and antibodies. Fibrous proteins are tough and water-insoluble. They are used to build a variety of materials that support and protect specific tissues. Example- Skin, Hair, Fingernails and Keratin. 

The sequence of amino acids in the chain is of critical importance in the biological functioning of the protein, and its determination is one of the most difficult problems in molecular biology. The chains may be relatively straight, or they may be coiled or helical. In the case of certain types of polypeptides, such as the keratins, they are cross-linked by the disulfide bonds of cystine. Linear polypeptides can be regarded as proteins. Synthesis of a 20-amino acid polypeptide that induces formation of antibodies for foot and mouth disease was announced in 1982. Itis the first vaccine to be synthesized. See protein polyamide keratin. 

Figure 4. Immunofluorescence microscopy of a cross-section of human skin exposed to saturated sulfur mustard vapor (1 min at 27 °C Ct 1040 mg.min.nr3 A) and of unexposed skin (B), using monoclonal antibody 1H10, directed against sulfur mustard adducts to human keratin, in a 1/50 dilution. The photographs are composed from an image obtained for FITC fluorescence (mainly emanating from the stratum corneum green) and from an image obtained for propidium iodide fluorescence representing DNA (red) in the same cross-section. (Reprinted with permission from G. R van der Schans et al., Chem. Res. Toxicol. 15, 21-25, 2002. Copyright (2002) American Chemical Society).

Antibodies against adducts of sulfur mustard to keratin in the skin are available which opens the way for the development of an assay for in situ detection of sulfur mustard exposure of human skin. 

Franko MC, Gibbs CJ, Rhoades DA, Gajdusek DC (1987) Monoclonal antibody analysis of keratin expression in the central nervous system. Proc. Natl Acad. Scl USA, 84, 3482-3485. 

---