kappa Light Chains Antikörper

Antigen: kappa Light Chains

Klonalität: Polyklonal

Wirt: Kaninchen

Reaktivität: Human

Applikation: Immunzytochemie (ICC), Immunpräzipitation (IP), Immunofixation (IFix), Western Blot (WB)

Zertifikate: ISO 9001:2008, ISO 13485:2003

Anbieter: Dako

Produktnummer: ABIN370524

Produktbeschreibung

Immunogen: Polyclonal immunoglobulin light chains of kappa type isolated from a pool of human sera.

Beschreibung: The human immunoglobulins basically consist of two identical heavy chains (Mr about 50 000) and two identical light chains (Mr about 20 000). The four chains are covalently linked together by disulphide bonds. The light chains are either kappa or lambda. The five immunoglobulins, IgA, IgD, IgE, IgG and IgM, differ regarding the heavy chains, and IgA and IgM also occur in polymeric forms (8). Individual B cells express either kappa or lambda light chains, but never both. In a polyclonal population the ratio of kappa to lambda bearing B cells is 2:1, and the occurrence of a mixture of kappa and lambda light chain-bearing cell types suggests polyclonality and a reactive or non-neoplastic proliferation of B cells (9). Diseases, such as multiple myeloma and B-cell lymphoma, are characterized by the proliferation of monoclonal neoplastic plasma cells producing only one type of light chain. Thus, the demonstration of a kappa or lambda light chain-restricted cell population is very useful in the histopathological diagnosis of these diseases (4). Primary AL (amyloid light-chain) amyloidosis is also a plasma cell disorder where the demonstration of monotypic kappa or lambda light chain in the amyloid deposits is diagnostically important and separates this disease from AA amyloidosis where immunoglobulin light chain deposits do not occur (3).

Spezifität: The antibody reacts with free kappa chains as well as kappa chains in intact immunoglobulin molecules. Traces of contaminating antibodies have been removed by solid-phase absorption with human plasma proteins. tests, staining with Coomassie Brilliant Blue: Double immunodiffusion: No reaction with lambda or gamma chains is seen when using 15 µ L of antibody against 15 µ L of lambda IgG. Crossed immunoelectrophoresis: Only kappa-related precipitates appear when using 12.5 µ L antibody per square cm gel area against 2 µ L human plasma or 2 µ L of a pool of concentrated urines from patients with tubular proteinuria. Cross-reaction with immunoglobulins from other species may occur. As demonstrated by immunocytochemistry, the antibody cross-reacts with immunoglobulins in dog, horse, pig and rat.

Anwendungen

Applikationshinweise: Polyclonal Rabbit Anti-Human Kappa Light Chains, code No. A 0191, is intended for use in immunocyto-chemistry. The antibody is also well-suited for gel immunoprecipitation techniques, including immunofixation and immunoblotting (1). In immunocytochemistry, the antibody labels plasma cells and related lymphoid cells containing kappa light chains, and it is a useful tool for the classification of patients with monoclonal gammopathies (2) and amyloidosis (3, 4). Additionally, the antibody may be used for distinguishing neoplastic monoclonal proliferation from reactive hyperplasia of B cells (5-7). Differential identification is aided by the results from 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 or B5 (2). Pre-treatment of tissues with proteinase K or heat- induced epitope retrieval is recommended. For heat-induced epitope retrieval, optimal results are obtained with DakoCytomation Target Retrieval Solution, code No. S 1700, DakoCytomation Target Retrieval Solution, High pH, code No. S 3308, 10 mmol/L citrate buffer, pH 6.0, or 10 mmol/L Tris buffer, 1 mmol/L EDTA, pH 9.0. The tissue sections should not dry out during the treatment or during the following immunocytochemical staining procedure. Frozen sections and cell preparations: The antibody can be used for labelling acetone-fixed, frozen sections (6). Dilution: Polyclonal Rabbit Anti-Human Kappa Light Chains, code No. A 0191, may be used at a dilution range of 1:1000-1:2000 when applied on formalin-fixed, paraffin-embedded sections of human tonsil and using 20 minutes heat-induced epitope retrieval in 10 mmol/L Tris buffer, 1 mmol/L EDTA, pH 9.0, and 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. Automation: The antibody is well-suited for immunocytochemical staining using automated platforms, such as the DakoCytomation Autostainer Performance characteristics: Cells labelled by the antibody display staining of the cell membrane and/or cytoplasm. Normal tissues: In normal tonsil, the antibody labels plasma cells, follicle centre cells and mantle zone cells providing a polyclonal pattern with a clear distinction between positive and negative cells (4).

Buffer: Purified immunoglobulin fraction of rabbit antiserum provided 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: Peterson, Brown, Crosson et al.: "Application of the immunoperoxidase technic to bone marrow trephine biopsies in the classification of patients with monoclonal gammopathies." in: American journal of clinical pathology, Vol. 85, Issue 6, pp. 688-93, 1986 (PubMed).

Taylor, Burns: "The demonstration of plasma cells and other immunoglobulin-containing cells in formalin-fixed, paraffin-embedded tissues using peroxidase-labelled antibody." in: Journal of clinical pathology, Vol. 27, Issue 1, pp. 14-20, 1974 (PubMed).

Becker: "Determination of antisera titres ing the single radial immunodiffusion method." in: Immunochemistry, Vol. 6, Issue 4, pp. 539-46 (PubMed).

Harris, Poppema, Data: "Demonstration of immunoglobulin in malignant lymphomas. Use of an immunoperoxidase technic on frozen sections." in: American journal of clinical pathology, Vol. 78, Issue 1, pp. 14-21, 1982 (PubMed).

Withold, Reinauer: "An immunoblotting procedure following agarose gel electrophoresis for detection of Bence Jones proteinuria compared with immunofixation and quantitative light chain determination." in: European journal of clinical chemistry and clinical biochemistry : journal of the Forum of European Clinical Chemistry Societies, Vol. 33, Issue 3, pp. 135-8, 1995 (PubMed).

Struom, Fogazzi, Banfi et al.: "Light chain deposition disease of the kidney. Morphological aspects in 24 patients." in: Virchows Archiv : an international journal of pathology, Vol. 425, Issue 3, pp. 271-80, 1995 (PubMed).

Marshall-Taylor, Cartun, Mandich et al.: "Immunohistochemical detection of immunoglobulin light chain expression in B-cell non-Hodgkin lymphomas using formalin-fixed, paraffin-embedded tissues and a heat-induced epitope retrieval technique." in: Applied immunohistochemistry & molecular morphology : AIMM / official publication of the Society for Applied Immunohistochemistry, Vol. 10, Issue 3, pp. 258-62, 2002 (PubMed).