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Monoclonal Mouse Anti-Human B-Cell-Specific Activator Protein Clone DAK-Pax5

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Monoclonal Mouse Anti-Human B-Cell-Specific Activator Protein Clone DAK-Pax5 Monoclonal Mouse Anti-Human B-Cell-Specific Activator Protein Clone DAK-Pax5 Codice M7307 Uso previsto Per uso diagnostico in vitro. Monoclonal Mouse Anti-Human B-Cell-Specific Activator Protein, Clone DAK-Pax5, è previsto per l’utilizzo in immunoistochimica. Gli anticorpi contro la proteina attivatrice specifica delle cellule B (BSAP) possono essere utili per identificare i linfociti B pro-, pre- e maturi e per classificare i linfomi (1 –4). Insieme a un gruppo di anticorpi, è particolarmente utile nell’identificazione differenziale del morbo di Hodgkin paragonato al linfoma a grandi cellule anaplastico di tipo T e delle cellule null (1, 3). L’interpretazione clinica di un’eventuale colorazione o della sua assenza deve essere integrata mediante studi morfologici avvalendosi di controlli adeguati e deve essere valutata nell’ambito dell’anamnesi del paziente e di altri test diagnostici da parte di un patologo qualificato. Sinonimi per BSAP, Pax5, NF-HB, S α-BP, NFS µ-B1, LR1 e EBB-1 (2, 4). l’antigene Riepilogo BSAP, nota anche come paired box protein 5 (Pax5), è un fattore di trascrizione espresso nelle cellule B. BSAP e spiegazioni appartiene alla famiglia dei geni PAX , che codifica i fattori di trascrizione coinvolti nello sviluppo delle cellule B. BSAP è espressa anche nei linfociti B pro-, pre- e maturi, ma non nelle cellule plasmatiche (3, 4). La scissione mirata del gene BSAP nei topi blocca lo sviluppo delle cellule B allo stadio pro-cellula B, suggerendo che la BSAP assuma un ruolo nel controllo dello sviluppo delle cellule B (2). Durante l’embriogenesi, l’espressione della BSAP è transitoriamente espressa nello sviluppo del sistema nervoso centrale. In seguito, l’espressione della BSAP è rilevata nel fegato fetale, dove è collegata alla comparsa di linfopoiesi B. Quindi, la BSAP potrebbe non solo avere un ruolo importante nello sviluppo delle cellule B, ma anche nello sviluppo neurale (1, 3, 5). In immunoistochimica, può essere difficile distinguere il linfoma di Hodgkin (CHL) dal linfoma a cellule grandi anaplastico di tipo T e delle cellule null (1). Nel CHL, le cellule di Hodgkin e Reed-Sternberg (celule HRS) esprimono BSAP, mentre le cellule HRS nella maggior parte dei casi non mostrano antigeni delle cellule B come CD19 o CD20 (6). La BSAP è un marker utile per la diagnosi del linfoma non Hodgkin delle cellule B e del linfoma di Hodgkin e dei tumori neuroendocrini (1-4, 6-8), nonostante la sua espressione nei sottogruppi dei tumori maligni epiteliali (7). Fare riferimento alle General Instructions for Immunohistochemical Staining della Dako o alle istruzioni sul sistema di rilevazione per le procedure IHC per: (1) Principio della procedura, (2) Materiali necessari ma non forniti, (3) Conservazione, (4) Preparazione dei campioni, (5) Procedura per la colorazione, (6) Controllo della qualità, (7) Risoluzione dei problemi, (8) Interpretazione della colorazione, (9) Limiti generali. Reagente fornito Anticorpo monoclonale murino fornito sotto forma di liquido sopranatante per colture cellulari dializzato contro tampone Tris/HCl 0,05 mol/L a pH 7,2 e sodio azide 0,015 mol/L. Clone: DAK-Pax5. Isotipo: IgG1, kappa. Concentrazione di IgG murine (mg/l): vedere l’etichetta stampata sulla fiala. La concentrazione della proteina tra i lotti potrebbe variare senza influenzare la diluizione ottimale. La titolazione di ogni lotto è confrontata e regolata in base a un lotto di riferimento, per garantire la prestazione della procedura di colorazione immunoistochimica da lotto a lotto. Immunogeno Peptide 17-merico sintetico dal dominio inibitore C-terminale della proteina. Specificità Nei test Western blotting del lisato delle cellule della milza murina, l’anticorpo marca una banda da 55 kDa corrispondente alla BSAP. Precauzioni 1. Per uso professionale. 2. Questo prodotto contiene sodio azide (NaN 3), una sostanza chimica fortemente tossica in forma pura. Sebbene non sia classificata come prodotto pericoloso, la sodio azide alle concentrazioni indicate può reagire con il rame e il piombo delle tubature formando azidi metalliche fortemente esplosive. Durante lo smaltimento, sciacquare le tubature con abbondante acqua per prevenire l’eventuale accumulo di azide metallica. 3. Adottare le normali procedure previste per il trattamento dei prodotti di origine biologica. 4. Indossare indumenti di protezione personale adeguati, per evitare il contatto con gli occhi e la pelle. 5. Smaltire la soluzione inutilizzata nel rispetto delle disposizioni locali, regionali e nazionali in materia. Conservazione Conservare a 2–8 °C. Non utilizzare dopo la data di scadenza stampata sulla fiala. Se i reagenti sono conservati in condizioni diverse da quelle specificate, le loro condizioni dovranno essere verificate dall’utente. Non sono stati osservati segni evidenti che suggeriscano l’instabilità di questo prodotto, pertanto è opportuno analizzare un controllo positivo e un controllo negativo insieme ai campioni dei pazienti. Qualora si ottenga una colorazione imprevista, che non sia cioè giustificata da un cambiamento delle procedure di laboratorio ma sia dovuta ad un probabile problema dell’anticorpo, contattare l’Assistenza Tecnica di Dako. (119102-001) M7307/IT/KDH/2008.10.07 p. 1/3 Dako Denmark A/S | Produktionsvej 42 | DK-2600 Glostrup | Denmark | Tel. +45 44 85 95 00 | Fax +45 44 85 95 95 | CVR No. 33 21 13 17 Specimen preparation Sezioni incluse in paraffina: (Allestimento tessuto) L’anticorpo può essere utilizzato per marcare sezioni di tessuto incluse in paraffina, fissate in formalina. compresi i materiali È necessario il pretrattamento di sezioni di tessuto deparaffinate con la tecnica del recupero termoindotto di epitopi necessari ma non (HIER). Si ottengono ottimi risultati pretrattando i tessuti con HIER utilizzando Dako EnVision™ FLEX Target forniti Retrieval Solution, Low pH (Codici K8005/K8015) oppure Dako Target Retrieval Solution (Codici S1700/S1699). Si consiglia il pretrattamento di sezioni di tessuto deparaffinate fissate in formalina, incluse in paraffina, utilizzando Dako PT Link (codici PT100/PT101). Per i dettagli, vedere la Guida Utente PT Link. Per PT Link, bisogna attenersi ai seguenti parametri: Temperatura di preriscaldamento: 65°C, temperatura e tempo di recupero dell’epitop o: 97 °C per 20 (±1) minuti; lasciare raffreddare a 65 °C. Rimuo vere ogni rack per vetrini dal serbatoio PT e immergere immediatamente i vetrini in un vaso/serbatoio (ad es. PT Link Rinse Station, codice PT109) contenente EnVision™ FLEX Wash Buffer (10X) diluito a temperatura ambiente (codice K8002/8012). Lasciare i vetrini nel Wash Buffer per 1–5 minuti. Le sezioni di tessuto non devono mai asciugarsi durante il pretrattamento o la procedura di colorazione immunoistochimica. Per una maggiore aderenza delle sezioni di tessuto ai vetrini, si consiglia l’utilizzo di Dako Silanized Slides (codice S3003). Sezioni congelate e strisci cellulari: L’anticorpo è consigliato per l’uso con per sezioni congelate, fissate in acetone oppure su strisci cellulari fissati. Procedura per la Diluizione: Monoclonal Mouse Anti-Human B-Cell-Specific Activator Protein, codice M7307, può essere utilizzato con colorazione compresi rapporti di diluizione di 1:15–1:30 quando lo si applica sulle sezioni pretrattate, fissate in formalina, incluse in i materiali necessari paraffina con un’incubazione di 20 minuti a temperatura ambiente. Si consiglia di diluire l’anticorpo in EnVision™ ma non forniti FLEX Antibody Diluent (codici K8006/K8016), in Dako Antibody Diluent (codice S0809) oppure Dako Real™ Antibody Diluent (codice S2022). Queste informazioni sono fornite esclusivamente come linee guida. La condizione ottimale può variare a seconda del tipo di campione e del metodo di preparazione adottato, pertanto dovrà essere definita autonomamente da ogni laboratorio. Il reagente di controllo negativo consigliato è Dako Negative Control, Mouse IgG1 (codice X0931), diluito alla stessa concentrazione di IgG1 dell’anticorpo primario. A meno che la stabilità dell’anticorpo e del controllo negativo diluiti non sia stata determinata nella procedura effettiva di colorazione, è opportuno diluire tali reagenti immediatamente prima dell’uso. I controlli positivi e negativi devono essere analizzati insieme ai campioni dei pazienti. I controlli positivi e negativi con i tessuti devono essere analizzati contemporaneamente, utilizzando lo stesso protocollo dei campioni dei pazienti. Il tessuto di controllo positivo deve comprendere cellule di tonsille e le cellule/strutture devono evidenziare gli schemi di reazione descritti per il tessuto in questione in “Caratteristiche prestazionali” in tutti i campioni positivi. Visualizzazione: EnVision FLEX+, Mouse, High pH, (codice K8002/K8012), che sostituisce il High pH Target Retrieval Solution di questo kit con EnVision FLEX Target Retrieval Solution, Low pH (10x), (codice K8005/K8015) utilizzando un’incubazione di 20 minuti a temperatura ambiente. Seguire la procedura fornita con i sistemi di visualizzazione selezionati. Automatizzazione: l’anticorpo è idoneo per le procedure di colorazione immunoistochimica su piattaforme automatizzate, come ad esempio Dako Autostainer, Autostainer Plus e Autostainer Link. Interpretazione Nelle cellule marcate dall’anticorpo è visibile una colorazione nucleare. dei risultati della colorazione Caratteristiche Tessuti normali: I tessuti linfoidi normali (linfonodo, milza e tonsilla) mostrano una colorazione della BSAP nucleare prestazionali forte nelle cellule B del centro follicolare e delle cellule B nella
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