alpha-Fetoprotein (AFP) Antikörper

Antigen: alpha-Fetoprotein (AFP)

Synonyme: FETA, HPAFP, AFP, MGC128267

Klonalität: Polyklonal

Wirt: Kaninchen

Reaktivität: Human

Applikation: Immunzytochemie (ICC), ELISA, Immunpräzipitation (IP)

Zertifikate: ISO 9001:2008, ISO 13485:2003

Anbieter: Dako

Produktnummer: ABIN370517

Produktbeschreibung

Weitere Bezeichnung: Alpha-1-Fetoprotein

Immunogen: Alpha-1-fetoprotein isolated from human cord serum.

Beschreibung: AFP is a 70 kDa glycoprotein containing 590 amino acid residues. Cells of the embryonic yolk sac, fetal liver, and fetal intestinal tract synthesize this glycoprotein (5, 6). AFP is a very useful marker for the detection of numerous developmental defects and various pathologies, whether cancerous or not (7). Expression of AFP has been demonstrated in about 44% of hepatocellular carcinomas, and in gonadal and extragonadal germ cells tumours, including yolk sac tumours. AFP has not been detected in normal adult tissue by immuno-staining (5). However, traces of AFP are found in normal adult sera and in greater concentrations in maternal and foetal sera and amniotic fluid (8).

Spezifität: The antibody reacts with human alpha-1-fetoprotein. Traces of contaminating antibodies have been removed by solid-phase absorption with human plasma proteins. The specificity of the antibody has been ascertained as follows: Crossed immunoelectrophoresis: Only the alpha-1-fetoprotein precipitation arch appears when using 12.5 µL A 0008 per square cm gel area against 2 µL of a human cord serum fraction. No precipitation arch is seen when the antibody is tested against 2 µL human plasma. Staining: Coomassie Brilliant Blue. ELISA: In indirect ELISA no significant reaction is observed with normal human plasma. As demonstrated by rocket immunoelectrophoresis, the antibody cross-reacts with the AFP-equivalent protein in cat and wildebeest (9).

Anwendungen

Applikationshinweise: Polyclonal Rabbit Anti-Human Alpha-1-Fetoprotein (AFP), code No. A 0008, is intended for the determination of human alpha-1-fetoprotein in immunocytochemistry (1, 2, 3), ELISA (4) and gel immunoprecipitation tech-niques. Positive results may aid in the identification of non-neoplastic and neoplastic liver disease, yolk sac tumours and mixed germ cell tumours. However, it is important that the antibody is not used alone, but together with a panel of antibodies. Interpretation of results must be made within the context of the patient's clinical history and other diagnostic tests by a certified professional. Introduction: The antibody can be used for labelling paraffin-embedded tissue sections fixed in formalin. Pre-treatment of tissues with proteinase K or heat-induced epitope retrieval is not required. The tissue sections should not dry out during the treatment or during the following immunocytochemical staining procedure. Dilution: Polyclonal Rabbit Anti-Human Alpha-1-Fetoprotein, code No. A 0008, may be used at a dilution range of 1:400-1:800 when applied on formalin-fixed, paraffin-embedded sections of human foetal liver, and using 30 minutes incubation at room temperature with the primary antibody. Optimal conditions may vary depending on specimen and preparation method, and should be determined by each individual laboratory. The recommended negative control is DakoCytomation Rabbit Immunoglobulin Fraction (Solid-Phase Absorbed), code No. X 0936, diluted to the same protein concentration as the primary antibody. Unless the stability in the actual test system has been established, it is recommended to dilute the product immediately before use or dilute in DakoCytomation Antibody Diluent, code No. S 0809. Visualization: DAKO LSAB/HRP kit, code No. K 0679, and DAKO EnVision/HRP kits, code Nos. K 4008 and K 4010, are recommended. Follow the procedure enclosed with the selected visualization kit. Performance characteristics: Cells labelled by the antibody display a cytoplasmic staining pattern (1). Normal tissues: In a 20-day old human conceptus, the antibody labels the endodermal cells of the inner surface of the yolk sac. In the amnion AFP is also demonstrated (10). Abnormal tissues: 189 orchidectomy specimens with germ cell tumours (95 pure seminomas and 94 non-seminomas) were studied with the antibody. All seminomas were AFP negative, while 66% of the non-seminomas were positively stained for AFP. 21% of pure embryonal carcinomas (EC), 25% of EC components in mixed tumours, all yolk sac tumour components, 20% of pure teratomas (T) and 47% of T components were AFP positive. Carcinoma-in-situ present in seminiferous tubules adjacent to the tumours was never positively stained for AFP (1). In stomach cancers with elevated serum AFP levels, 19/35 (54%) were labelled by the antibody (2). For double antibody sandwich ELISA of alpha-1-fetoprotein, DakoCytomation General ELISA Procedure (Order No. 30 023) may serve as a guide. The procedure below is for the measurement of AFP in serum (range 5-200 µg/L). 1. Coating antibody: DakoCytomation Rabbit Anti-Human Alpha-1-Fetoprotein, code No. A 0008, diluted to a protein concentration of 10 mg/L. 2. Standard: DakoCytomation Alpha-1-Fetoprotein Standard, code No. X 0900, diluted 1:2 750-1:110 000. Dilution of test samples (serum): 1:5. 3. Detection antibody: DakoCytomation Peroxidase-Conjugated Rabbit Anti-Human Alpha-1-Fetoprotein, code No. P 0128. Guideline for dilution 1:3 000. Unless the stability in the actual test system has been established, it is recommended to dilute the product immediately before use.

Buffer: Purified immunoglobulin fraction of rabbit antiserum in liquid form. In 0.1 mol/L NaCl, 15 mmol/L NaN 3 .

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, sodium azide 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. 3. As with any product derived from biological sources, proper handling procedures should be used. 4. The product may be used in different techniques and in combination with different sample types and materials, therefore each individual laboratory should validate the test system applied.

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

Publikationen

Publikationen: MacDonald, Kelly: "The rapid quantitation of serum alpha-fetoprotein by two-site micro enzyme immunoassay." in: Clinica chimica acta; international journal of clinical chemistry, Vol. 87, Issue 3, pp. 367-72, 1978 (PubMed).

Perrone, Steeper, Dehner: "Alpha-fetoprotein localization in pure ovarian teratoma. An immunohistochemical study of 12 cases." in: American journal of clinical pathology, Vol. 88, Issue 6, pp. 713-7, 1987 (PubMed).

Kodama, Kameya, Hirota et al.: "Production of alpha-fetoprotein, normal serum proteins, and human chorionic gonadotropin in stomach cancer: histologic and immunohistochemical analyses of 35 cases." in: Cancer, Vol. 48, Issue 7, pp. 1647-55, 1981 (PubMed).

Jacobsen, Jacobsen: "Alpha-fetoprotein (AFP) and human chorionic gonadotropin (HCG) in testicular germ cell tumours. A prospective immunohistochemical study." in: Acta pathologica, microbiologica, et immunologica Scandinavica. Section A, Pathology, Vol. 91, Issue 3, pp. 165-76, 1983 (PubMed).

Jacobsen, Jacobsen, Henriksen: "An immunohistochemical study of a series of plasma proteins in the early human conceptus." in: Oncodevelopmental biology and medicine : the journal of the International Society for Oncodevelopmental Biology and Medicine, Vol. 2, Issue 6, pp. 399-410, 1982 (PubMed).

Nunez: "Biological role of alpha-fetoprotein in the endocrinological field: data and hypotheses." in: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine, Vol. 15, Issue 2, pp. 63-72, 1994 (PubMed).

Mizejewski: "alpha-fetoprotein as a biologic response modifier: relevance to domain and subdomain structure." in: Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.), Vol. 215, Issue 4, pp. 333-62, 1997 (PubMed).