Actin, Muscle Antikörper

Antigen: Actin, Muscle

Klonalität: Monoklonal (HHF35)

Wirt: Maus

Reaktivität: Human

Applikation: Immunhistochemie (IHC), Immunhistochemie (IHC)

Zertifikate: ISO 9001:2008, ISO 13485:2003

Anbieter: Dako

Produktnummer: ABIN370558

Produktbeschreibung

Weitere Bezeichnung: Muscle Actin

Immunogen: SDS extracted protein fraction of human myocardium

Isotyp: IgG1, kappa chain  (Passende Sekundärantikörper)

Klon: HHF35

Beschreibung: Actin, a highly conserved, ubiquitous cytoskeletal protein of muscle and nonmuscle cells, exists in three isotypes ( alpha , beta , gamma ) that differ by their amino acid sequences and isoelectric points. The monoclonal mouse anti-human Muscle Actin, clone HHF35 was made by immunizing mice with a polypeptide fraction of human myocardium from a case of idiopathic hypertrophic subaortic stenosis.

Spezifität: Actin does not react with the alpha -actin of non-muscle (endothelial cells) sources. 1 Gel electrophoresis and immunoblots show the specificity of HHF35 to be for the alpha - and gamma -actin isotypes of skeletal, cardiac and smooth muscle. 1

Anwendungen

Applikationshinweise: This antibody is intended for laboratory use to identify qualitatively by light microscopy an epitope present on muscle actin in normal and neoplastic tissues using immunohistochemical (IHC) test methods. Paraffin Sections Anti-Muscle Actin, HHF35 can be used on formalin-fixed, paraffin-embedded tissue sections. Pretreatment of tissue with proteolytic enzymes is not required. Cryostat Sections and Cell Smears Anti-Muscle Actin, HHF35 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 anti-muscle actin is cytoplasmic. Product specific limitations 1. Schmidt, et al. and others found that the addition of EDTA to an HHF35 primary antibody diluent reduced nonspecific staining 2,3 and also decreased the chances of false-positive staining of neuroblastomas, retinoblastomas, and Ewing Sarcomas while maintaining adequate sensitivity for myogenic tumors. 3 2. Miettinen found mild enzyme predigestion (pepsin, pronase, trypsin) to improve staining quality of formalin-fixed, paraffin-embedded tissue, 7 however, Dako does not recommend tissue pretreatment. 3. Only rarely was immunoreactivity with HHF35 observed in isolated spindle cells of the liver, lymph nodes, kidney, pancreas, and the adrenal gland. 8 4. Neoplastic cells of some pleomorphic undifferentiated sarcomas (malignant fibrous histiocytomas, MFH) have been reported positive, localized only to the smooth muscle cells and pericytes of blood vessels. 2,7 Normal Tissues In normal tissue, HHF35 demonstrates cytoplasmic staining of striated fibers of skeletal muscles, the smooth muscles of arteries, veins and pericytes of smaller arteries, the tunica muscularis of the GI tract, the myometrium of the uterus, prostatic stroma, the capsule cells of several parenchymal organs, including liver, kidney, lymph nodes and spleen, and the myoepithelial layers of the mammary ducts and glands, and the eccrine sweat, bronchial and salivary glands. 1,2,7-9 Other non-muscle cells are non-reactive, including vascular endothelial cells, epithelial cells, lymphoid cells, macrophages, connective tissue, and neural cells. 1,2,8,9 Abnormal Tissues In pathological tissues, HHF35 was demonstrated to be a reliable marker for soft tissue tumors with muscle differentiation, i.e. leiomyomas (LM), leiomyosarcomas (LMS) and rhabdomyosarcomas (RMS), for which it displayed a higher degree of sensitivity than desmin antibodies. 2 This was confirmed by Schmidt, et al. 3 who found 29/30 RMS, including embryonal, alveolar, botryoid and pleomorphic subtypes, regardless of the degree of differentiation, to be HHF35 positive. A study comprising 285 well characterized soft tissue tumors 7 found 17/17 RMS, 31/32 LMS, 23/23 LM and 3/5 pleomorphic liposarcomas to be immunoreactive with HHF35. The majority of glomus tumors also reacted with HHF35. 7,10 Desmoid tumors showed occasional positive cells in 9/15 cases. 7 Similar results were reported by others 8 who found 34/35 RMS, 11/22 LMS, 5/6 LM and 4/4 rhabdomyomas to be HHF35 reactive. The myofibroblasts of some lesions, including reactive tissue, healing wounds and atherosclerotic plaques also stained with HHF35 in the majority of cases. 1,2,8,11 HHF35 was also used successfully for the differentiation of noninvasive (consistently actin positive) from invasive breast tumors (actin negative). 9 Non-muscle sarcomas and neoplastic cells of carcinomas, melanomas, and lymphomas are non-reactive.

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. 4,5 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.

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

Bilder

Leiomyosarcoma. The majority of the neoplastic cells show amoderate to strong staining reaction.

Rhabdomyosarcoma. The majority of the neoplastic cellsshow a weak to moderate staining reaction.

Colon. All the smooth muscle cells in vessel walls, lamina muscularis mucosa and myoepithelial cells around colonic crypts show a moderate to strong staining reaction.

Publikationen

Publikationen: Kivelae, Fuchs, Tarkkanen: "Cytoskeleton in neuroectodermally derived epithelial and muscle cells of the human iris and ciliary body." in: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, Vol. 40, Issue 10, pp. 1517-26, 1992 (PubMed).

Bohn, Wiegers, Beuttenmueller et al.: "Species-specific recognition patterns of monoclonal antibodies directed against vimentin." in: Experimental cell research, Vol. 201, Issue 1, pp. 1-7, 1992 (PubMed).

Azumi, Battifora: "The distribution of vimentin and keratin in epithelial and nonepithelial neoplasms. A comprehensive immunohistochemical study on formalin- and alcohol-fixed tumors." in: American journal of clinical pathology, Vol. 88, Issue 3, pp. 286-96, 1987 (PubMed).

Osborn, Debus, Weber: "Monoclonal antibodies specific for vimentin." in: European journal of cell biology, Vol. 34, Issue 1, pp. 137-43, 1984 (PubMed).

Herrmann, Aebi: "Intermediate filaments and their associates: multi-talented structural elements specifying cytoarchitecture and cytodynamics." in: Current opinion in cell biology, Vol. 12, Issue 1, pp. 79-90, 2000 (PubMed).