Cytokeratin 1 (KRT1) Antikörper

Antigen: Cytokeratin 1 (KRT1)

Synonyme: K1, CK1, EHK, EHK1, EPPK, KRT1A, NEPPK

Klonalität: Monoklonal (AE1-AE3 1-2)

Wirt: Maus

Reaktivität: Human

Applikation: Immunhistochemie (Paraffinschnitte) (IHC (p))

Zertifikate: ISO 9001:2008, ISO 13485:2003

Anbieter: Dako

Produktnummer: ABIN370633

Produktbeschreibung

Weitere Bezeichnung: Cytokeratin

Immunogen: Human epidermal callus 1

Isotyp: IgG1, kappa chain  (Passende Sekundärantikörper)

Klon: AE1-AE3 1-2

Beschreibung: Cytokeratins are a family of water-soluble proteins with molecular weights between 40-70 kD that form the cytoskeleton of epithelial cells. At least 19 different cytokeratins have been identified and can be divided into two subfamilies. Subfamily A comprises relatively acidic cytokeratins (with a pI under 5.5) whereas members of subfamily B have a relatively basic pI of 6 or over.

Spezifität: AE1/AE3 is a cocktail of two monoclonal antibodies that were obtained by immunizing mice with human callus keratins. 2 AE1/AE3 has been shown to identify the majority of human cytokeratins and thus may be used as a tool for the positive IHC identification of cells of simple and stratified epithelial origin. 1,2,4 Antibody AE1 immunoreacts with an antigenic determinant present on most of the subfamily A cytokeratins, including cytokeratins with Molls designation 4 10, 13, 14, 15 16 and 19 (MWs of 56.5, 54, 50, 50, 48 and 40 kD, respectively) but not on Nos. 12, 17 and 18 (55, 47 and 45 kD). 4 Antibody AE3 reacts with an antigenic determinant shared by the subfamily B cytokeratins including Nos. 1 and 2, 3, 4, 5, 6, 7 and 8 (MWs of 65, 67, 64, 59, 58, 56, 54 and 52 kD, respectively). 5

Anwendungen

Applikationshinweise: Monoclonal mouse anti-human Cytokeratin, clones AE1/AE3 is intended for laboratory use to identify qualitatively by light microscopy two epitopes present on a majority of epithelial cytokeratins in acetone-fixed frozen and formalin-fixed, paraffin-embedded tissue using immunohistochemical (IHC) test methods. Positive results aid in the classification of normal and neoplastic tissue as epithelial in origin 1-3 and serve as an adjunct to conventional histopathology. The clinical interpretation of any positive staining or its absence should be complemented by morphological and histological studies with proper controls. Evaluations should be made within the context of the patient's clinical history and other diagnostic tests by a qualified individual. Paraffin Sections AE1/AE3 can be used on formalin-fixed, paraffin-embedded tissue sections. Tissue pretreatment for epitope enhancement is required. Either digestion using proteolytic enzymes or heat-induced epitope retrieval is effective. The following enzymes can be used for pretreatment of formalin-fixed, paraffin-embedded tissues: Proteinase K, RTU (code S3020), Pepsin (code S3002), Pronase (code S2013), Trypsin (code S2012) or Proteolytic Enzyme, RTU (code S3007). Rinse thoroughly with distilled water and continue with the staining procedure of the detection system instructions. As an alternative to enzyme pretreatment, heat-induced epitope retrieval can be used. Heat-induced epitope retrieval involves immersion of tissue sections in a pre-heated buffer solution and maintaining heat, either in a water bath (95-99 °C ), a steamer (95-99 °C) or an autoclave (121 °C). For greater adherence of tissue sections to glass slides, the use of Silanized Slides (code S3003) is recommended. Target Retrieval Solution (code S1700) or 10x Concentrate (code S1699) is recommended using a 20-minute heating protocol. After thermal treatment, allow the jar with buffer and slides to cool for 20 minutes at room temperature. Rinse well with buffer. Cryostat Sections and Cell Smears AE1/AE3 can be used for labelling acetone-fixed cryostat sections or fixed cell smears. Follow the procedure for the detection system selected. Staining interpretation The cellular staining pattern for AE1/AE3 is cytoplasmic. Product specific limitations 1. Extrafollicular reticulum cells of lymph nodes, tonsil and spleen have been shown to react with antibodies to cytokeratin 8. 7 2. The presence of cytokeratin 19 and possibly 8 has been confirmed in smooth muscle cells of the uterus. 8 3. Rare melanomas 9 and leiomyosarcomas 8 may stain positive. This finding is usually more pronounced on frozen tissue rather than formalin-fixed tissue. 10 Therefore, it is recommended that AE1/AE3 be used in a panel with HMB45 (when ruling out melanomas) and desmin (when ruling out leiomyosarcomas) as these latter antibodies lack specificity for epithelial cells. 4. False-positive staining of glial cells in tumors has been reported on formalin-fixed, paraffin-embedded tissue when proteolytic pretreatment was employed. It was shown by immunocytochemical and biochemical methods that these cells and tumors do not express cytokeratins. 11 5. Pinkus et al. 12 stressed the importance of proteolytic digestion of formalin-fixed tissues to be stained with AE1/AE3. Photos of stained tissues depicted include the results when digestion with trypsin II was omitted, other trypsin enzymes were used and/or when suboptimal digestion techniques were applied. In the latter cases, only 2/12 epithelial neoplasms of various types exhibited optimal staining for cytokeratin. By reviewing conflicting cytokeratin immunoreactivities in earlier publications and comparing the same with their own, Pinkus et al. 12 considered many previously false-negative cases attributable to one or several of these short-comings. 6. From a comparison of AE1/AE3 with an anti-epithelial membrane antigen (EMA) antibody in an IHC study of 87 neoplasms, including 48 adenocarcinomas of various types, Pinkus et al. 13 concluded that the cytokeratin proteins in proteolytically treated formalin-fixed tissues and as stained by AE1/AE3 were more reliable markers in 33% of the cases of epithelial derived neoplasms than anti-EMA. However, because EMA was positive and AE1/AE3 negative in 9% of the cases, it was recommended that AE1/AE3 and anti-EMA be used as complementary reagents. 7. Although no false-positive staining was reported by Listrom and Dalton, 14 faint cytoplasmic staining was observed in 2/2 plasmacytomas, 2/4 melanomas and 2/7 lymphomas and considered to be the result of nonspecific background staining. Normal Tissues Testing of 30 different normal tissues demonstrated positive staining in the cytoplasm of squamous and columnar epithelium of the cervix, colon, esophagus, skin, small intestine, stomach and tonsil. Other tissues that stained included glandular tissue (mammary, parathyroid, prostate sweat and thyroid), astrocyte, white matter of the cerebellum, glial filaments of the cerebrum, distal tubule and Bowman's capsule of the kidney, bile duct, pneumocytes, bronchi, mesothelium, interlobular duct of the pancreas, anterior pituitary cell, interlobular duct and acinar cells of the salivary gland, reticular cells and Hassall's bodies of the thymus, and endometrium and smooth muscle of the uterus. 15 Negative staining was noted for adrenal, bone marrow, heart, pericardium, peripheral nerve, skeletal muscle, spleen and testis. AE1/AE3 reacts with keratinized (56.5/65-67) and corneal (55/64) epidermis, stratified squamous epithelia of internal organs (51/59), stratified epithelia (50/58), hyperproliferative keratinocytes (48/56) and simple epithelia (45/52 and 46/54). The 40 kD keratin is present in most epithelia except adult epidermis. 3,4 Abnormal Tissues In pathological tissues, Listrom and Dalton 14 tested clones AE1/AE3 on over 60 poorly differentiated epithelial neoplasms, lymphomas, melanomas and sarcomas. Except for staining of only 2/6 cases of small cell carcinoma and 3/5 transitional cell carcinoma, the study found all of 34 epithelial neoplasms to stain positively. When positive, AE1/AE3 stained transitional cell carcinomas only weakly and staining of the tumor cells was either diffusely cytoplasmic or perinuclear. Montag et al. 16 found AE1/AE3 to be a sensitive reagent for the differential diagnosis of diffuse malignant mesothelioma of the sarcomatoid (spindle-cell) type (positive in 30/30 cases) from other types of spindle cell neoplasms (0/49). When compared with anti-EMA in a study of 87 neoplasms, including 48 adenocarcinomas of various types, Pinkus et al. 13 found AE1/AE3 to stain 33% of the cases more reliably than the anti-EMA. Although 3/3 cases of chondroid chordoma and 1/8 cases of lymphoma were reactive with anti-AE1/AE3, no positive staining was observed among 25 non-epithelial neoplasms including 4 cases each of melanoma and glioblastoma. 14 No cytokeratin was detected in 8 cases of nonepithelial tumors, including melanoma, lymphoma, neurofibroma, and sarcoma when AE1/AE3 was used in the immunoblot technique. 17 However, it was recommended 14 that AE1/AE3 serve as a member of an antibody panel for the determination of cell lineage in cases of poorly differentiated small cell neoplasms. (105470-004) 303322EFG_002 p.

Buffer: Monoclonal Mouse antibody provided in liquid form as tissue culture supernatant in 0.05 mol/L Tris-HCl, pH 7.2 and 0.015 mol/L sodium azide. This product contains stabilizing protein.

Lagerung: Store at 2-8 °C. Precautions: 1. For professional users. 2. This product contains sodium azide (NaN 3 ), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, NaN 3 may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing. 6 3. As with any product derived from biological sources, proper handling procedures should be used. 4. Wear appropriate Personal Protective Equipment to avoid contact with eyes and skin. 5. Unused reagents should be disposed of according to local, State, and Federal regulations.

Forschungsgebiet: Extrazelluläre Matrix

Beschränkungen: Für Forschungszwecke. Als CE-zertifizierter Antikörper in der Europäischen Union für In Vitro Diagnostik (IVD) zugelassen.

Publikationen

Publikationen: Groome, Hancock, Betteridge et al.: "Monoclonal and polyclonal antibodies reactive with the 1-32 amino terminal sequence of the alpha subunit of human 32K inhibin." in: Hybridoma, Vol. 9, Issue 1, pp. 31-42, 1990 (PubMed).

Costa, Ames, Walls et al.: "Inhibin immunohistochemistry applied to ovarian neoplasms: a novel, effective, diagnostic tool." in: Human pathology, Vol. 28, Issue 11, pp. 1247-54, 1997 (PubMed).

Matias-Guiu, Pons, Prat: "Müllerian inhibiting substance, alpha-inhibin, and CD99 expression in sex cord-stromal tumors and endometrioid ovarian carcinomas resembling sex cord-stromal tumors." in: Human pathology, Vol. 29, Issue 8, pp. 840-5, 1998 (PubMed).

Arola, Liu, Heikkilae et al.: "Expression of inhibin alpha in the human adrenal gland and adrenocortical tumors." in: Endocrine research, Vol. 24, Issue 3-4, pp. 865-7, 1999 (PubMed).

La Rosa, Uccella, Billo et al.: "Immunohistochemical localization of alpha- and betaA-subunits of inhibin/activin in human normal endocrine cells and related tumors of the digestive system." in: Virchows Archiv : an international journal of pathology, Vol. 434, Issue 1, pp. 29-36, 1999 (PubMed).

Di Loreto, Reis, Cataldi et al.: "Human mammary gland and breast carcinoma contain immunoreactive inhibin/activin subunits: evidence for a secretion into cystic fluid." in: European journal of endocrinology / European Federation of Endocrine Societies, Vol. 141, Issue 2, pp. 190-4, 1999 (PubMed).

Huang, Gao, Boini et al.: "In vivo stimulation of AMP-activated protein kinase enhanced tubuloglomerular feedback but reduced tubular sodium transport during high dietary NaCl intake." in: Pflügers Archiv : European journal of physiology, 2010 (PubMed).