EnVision FLEX, High PH (Dako Omnis)
EnVision FLEX, High pH (Dako Omnis)Code GV8002nd edition/ 2ème édition / 2. AuflageThe kit contains reagents sufficient for 600 tests. For Dako Omnis instruments.Les réactifs contenus dans ce kit permettent d’effectuer 600 tests. Pour les appareils Dako Omnis.Die in diesem Kit enthaltenen Reagenzien sind ausreichend für 600 Tests. Für Dako Omnis Geräte.Optional reagents/ Réactifs en option/ Optionale Reagenzien:CodeProduct name/ Nom du produit/ ProduktnameTestsGV804EnVision FLEX Target Retrieval Solution, High pH (50x)(Dako Omnis)225GV805EnVision FLEX Target Retrieval Solution, Low pH (50x)(Dako Omnis)225GC807Wash Buffer (20x) (Dako Omnis)GC808Hematoxylin (Dako Omnis)600GV809EnVision FLEX Rabbit LINKER (Dako Omnis)75GV821EnVision FLEX Mouse LINKER (Dako Omnis)75GV825EnVision FLEX DAB Substrate Chromogen System(Dako Omnis)150Other options/ Autres options/ Weitere Optionen:K8020(125835-001)FLEX IHC Microscope Slides500P04120EFG 01 GV800/2015.05 p. 1/32
Contents/ Table des matières/ InhaltPage/ Page/ SeiteENGLISHIntended Use . 4Summary and Explanation. 4Reagents . 5A. Materials provided . 5B. Materials required but not provided. 5C. Optional reagents . 6Precautions . 7Storage. 8Reagent Preparation. 9Specimen Collection and Preparation . 9Procedure. 10Staining procedure . 10Quality Control. 11Interpretation of Results. 11Limitations . 12Explanation of symbols. 32FRANÇAISUtilisation prévue . 13Résumé et explication . 13Réactifs . 14A. Matériel fourni. 14B. Matériel requis mais non fourni . 14C. Réactifs en option. 15Précautions d'emploi . 16Conservation . 17Préparation des réactifs. 18Prélèvement et préparation des échantillons . 19Procédure. 19Procédure de coloration . 19Contrôle qualité . 22Interprétation des résultats . 22Limites . 22Explication des symboles. 32(125835-001)P04120EFG 01 GV800/2015.05 p. 2/32
DEUTSCHVerwendungszweck. 23Zusammenfassung und Erklärung . 23Reagents (Reagenzien) . 24A. Mitgelieferte Materialien. 24B. Erforderliches, aber nicht mitgeliefertes Material. 24C. Optionale Reagenzien . 25Hinweise und Vorsichtsmaßnahmen. 26Lagerung . 27Reagenzienvorbereitung. 28Entnahme und Vorbereitung der Probe. 28Verfahren. 29Färbeverfahren . 29Qualitätskontrolle. 31Auswertung der Ergebnisse. 31Einschränkungen. 31Erläuterung der Symbole . 32(125835-001)P04120EFG 01 GV800/2015.05 p. 3/32
ENGLISHIntended UseFor in vitro diagnostic use.EnVision FLEX, High pH (Dako Omnis) visualization system is intended for use inimmunohistochemistry together with Dako Omnis instruments. The system detects primary mouse andrabbit antibodies and the reaction is visualized by EnVision FLEX DAB Chromogen. If used withEnVision FLEX Rabbit LINKER (Code GV809) or EnVision FLEX Mouse LINKER (Code GV821)signal amplification of primary rabbit antibodies or primary mouse antibodies, respectively, can beachieved. The EnVision FLEX reagents are intended for use on formalin-fixed, paraffin-embeddedtissue sections.Summary and ExplanationEnVision FLEX visualization system is a flexible system giving optimal staining on Dako Omnisinstruments, when using the protocol recommended in this package insert.Prior to staining formalin-fixed, paraffin-embedded tissue sections are subjected to onboarddeparaffinization and hydration followed by heat-induced epitope retrieval (HIER) using the targetretrieval reagent specified in the package insert for the primary antibody. This kit includes EnVision FLEX Target Retrieval Solution, High pH (50x) (DM848). Alternatively, EnVision FLEX TargetRetrieval Solution, High pH (50x) (Code GV804) or EnVision FLEX Target Retrieval Solution, Low pH(50x) (Code GV805) is recommended. Some primary antibodies require enzymatic pre-treatment oftissue for optimal staining instead of HIER.Endogenous peroxidase should be blocked with EnVision FLEX Peroxidase-Blocking Reagent(DM841) included in the kit. Due to an effective washing procedure and the presence of stabilizingproteins in the Dako reagents, extra blocking steps to reduce non-specific background staining areunnecessary.Wash Buffer (20x) (GC807) available as optional reagent is recommended.Primary antibodies are not provided with the kit. We recommend the use of Dako FLEX Ready-to-UsePrimary Antibodies or Dako concentrated Primary Antibodies (see respective package inserts forguidelines).EnVision FLEX Rabbit LINKER (Code GV809) may be applied for an optional signal amplification ofrabbit primary antibodies. For signal amplification when using mouse primary antibodies, EnVision FLEX Mouse LINKER (Code GV821) may be applied. As a guideline: EnVision FLEX RabbitLINKER provides 2-3 fold signal amplification and EnVision FLEX Mouse LINKER provides 4-5 foldsignal amplification.Dako EnVision FLEX /HRP reagent (DM842) in the kit consists of a dextran backbone to which alarge number of horseradish peroxidase (HRP) molecules and secondary antibody molecules havebeen coupled. A unique chemistry is used for the coupling reaction, which permits the binding of up to100 HRP molecules and up to 20 antibody molecules per backbone.The substrate system in the kit consists of two components: EnVision FLEX DAB Chromogen(DM847), a concentrated diaminobenzidine (DAB) solution, and EnVision FLEX Substrate Buffer(DM843) containing hydrogen peroxide. Before use EnVision FLEX DAB Chromogen must bediluted in EnVision FLEX Substrate Buffer. The substrate system produces a crisp brown endproduct at the site of the target antigen. Timely mixing of chromogen and buffer is automaticallyperformed onboard Dako Omnis.Hematoxylin (Code GC808) is recommended for counterstaining. The reagent provides a clear blue,nuclear staining.The stained tissue sections should be mounted with organic-solvent-based mounting medium.(125835-001)P04120EFG 01 GV800/2015.05 p. 4/32
ReagentsA. Materials providedDM841EnVision FLEX Peroxidase-Blocking Reagent (Dako Omnis)8 x 22.5 mL, ready-to-usePhosphate buffer containing hydrogen peroxide, 15 mmol/L NaN3 anddetergent.DM842EnVision FLEX /HRP (Dako Omnis)8 x 22.5 mL, ready-to-useDextran coupled with peroxidase molecules and goat secondary antibodymolecules against rabbit and mouse immunoglobulins. In buffered solutioncontaining stabilizing protein and preservative.DM847EnVision FLEX DAB Chromogen (Dako Omnis)8 x 1 mL3,3’-diaminobenzidine tetrahydrochloride in organic solvent.The color of this reagent may vary from strong violet to colorless withouthaving any influence on the performance of the kit.DM843EnVision FLEX Substrate Buffer (Dako Omnis)16 x 26 mLBuffered solution containing hydrogen peroxide and preservative.DM848EnVision FLEX Target Retrieval Solution, High pH (50x) (Dako Omnis)9 x 68 mL, 50x concentratedTris/EDTA buffer, pH 9B. Materials required but not providedAntibody diluent, e.g. Dako Antibody Diluent (Code S0809)Clearify Clearing Agent (Code GC810)Dako FLEX Ready-to-Use Primary Antibodies or suitably diluted concentrated primary rabbit or mouseantibodies from DakoDako Omnis instrumentDako Omnis instrumentation utensilsDako Proteinase K, Ready-to-Use (if necessary) (Code S3020)FLEX Universal Negative Control, Rabbit, Ready-to-Use (Code GA600), FLEX Universal NegativeControl, Mouse, Ready-to-Use (Code GA750), or suitable negative control reagent for the primaryantibodyHematoxylin (Dako Omnis) (Code GC808) or suitable hematoxylin reagent for counterstainingMicroscope slides, e.g. Dako FLEX IHC Microscope Slides (Code K8020)Mounting medium (organic-solvent-based) and coverslipsWash Buffer (20x) (Dako Omnis) (Code GC807)0.3 M Sulfuric Acid (Code GC203)(125835-001)P04120EFG 01 GV800/2015.05 p. 5/32
C. Optional reagentsGV804DM848EnVision FLEX Target Retrieval Solution, High pH (50x) (Dako Omnis)3 x 68 mL, 50x concentratedTris/EDTA buffer, pH 9GV805DM849EnVision FLEX Target Retrieval Solution, Low pH (50x) (Dako Omnis)3 x 68 mL, 50x concentratedCitrate buffer, pH 6.1GC807DM851Wash Buffer (20x) (Dako Omnis)20 x 175 mL, 20x concentratedTris-buffered saline solution containing Tween 20, pH 7.6 ( 0.1).GC808DM846Hematoxylin (Dako Omnis)8 x 22.5 mL, ready-to-useAqueous solution of hematoxylin.GV809DM845EnVision FLEX Rabbit LINKER (Dako Omnis)1 x 22.5 mL, ready-to-useBuffered solution containing stabilizing protein and an antimicrobial agent.GV821DM844EnVision FLEX Mouse LINKER (Dako Omnis)1 x 22.5 mL, ready-to-useBuffered solution containing stabilizing protein and an antimicrobial agent.GV825DM847 DM843EnVision FLEX DAB Substrate Chromogen System (Dako Omnis)EnVision FLEX DAB Chromogen (Dako Omnis)2 x 1 mL3,3’-diaminobenzidine tetrahydrochloride in organic solvent.The color of this reagent may vary from strong violet to colorless without havingany influence on the performance of the kit.EnVision FLEX Substrate Buffer (Dako Omnis)4 x 26 mLBuffered solution containing hydrogen peroxide and preservative.(125835-001)P04120EFG 01 GV800/2015.05 p. 6/32
Precautions1.2.3.4.For professional users.EnVision FLEX /HRP contains material of animal origin and it cannot be excluded that traceamounts of human material could be present due to manufacturing procedures. As with anyproduct derived from biological sources, proper handling should be used.Do not expose EnVision FLEX Peroxidase-Blocking Reagent, EnVision FLEX /HRP,EnVision FLEX Substrate Buffer, EnVision FLEX DAB Chromogen or EnVision FLEXSubstrate Working Solution to strong light.EnVision FLEX Target Retrieval Solution, High pH (50x) contains 5-10% trometamol, 1-5%nonylphenol, ethoxylated, and 1-5% acetic acid, (ethylenedinitrilo)tetra-, disodium salt, dihydrate.EnVision FLEX Target Retrieval Solution, High pH (50x) is labeled:WarningH319H411P280P273P264P305 P351 P3385.6.Causes serious eye irritation.Toxic to aquatic life with long lasting effects.Wear eye or face protection.Avoid release to the environment.Wash hands thoroughly after handling.IF IN EYES: Rinse cautiously with water for several minutes.Remove contact lenses, if present and easy to do. Continue rinsing.P337 P313If eye irritation persists: Get medical attention.P501Dispose of contents and container in accordance with all local,regional, national and international regulations.For EnVision FLEX Peroxidase-Blocking Reagent, a safety data sheet is available forprofessional users on request.EnVision FLEX DAB Chromogen contains 1-5% de, and is labeled:DangerH350H341P201P2027.8.May cause cancer.Suspected of causing genetic defects.Obtain special instructions before use.Do not handle until all safety precautions have been read andunderstood.P280Wear protective gloves. Wear eye or face protection. Wear protectiveclothing.P308 P313IF exposed or concerned: Get medical attention.P405Store locked up.P501Dispose of contents and container in accordance with all local,regional, national and international regulations.As a general rule, persons under 18 years of age are not allowed to work with this product.Users must be carefully instructed in the proper work procedure, the dangerous properties of theproduct and the necessary safety instructions. Please refer to the Safety Data Sheet for additionalinformation.Wear appropriate Personal Protective Equipment to avoid contact with eyes and skin.Unused solution should be disposed of according to local, State and Federal regulations.(125835-001)P04120EFG 01 GV800/2015.05 p. 7/32
StorageStore EnVision FLEX Target Retrieval Solution, High pH (50x) at 2-8 C. Store EnVision FLEXPeroxidase-Blocking Reagent, EnVision FLEX /HRP, EnVision FLEX DAB Chromogen andEnVision FLEX Substrate Buffer in the dark at 2-8 C. During storage the cap should be closed.Onboard stability of EnVision FLEX Peroxidase-Blocking Reagent, EnVision FLEX /HRP,EnVision FLEX DAB Chromogen and EnVision FLEX Substrate Buffer is 75 hours. Onboardstability of diluted EnVision FLEX Target Retrieval Solution, High pH and Low pH is 7 days.Remaining onboard stability is tracked by the Dako Omnis software.Do not use after expiration date stamped on the vials. If reagents are stored under any conditions otherthan those specified, the conditions must be verified by the user. If unexpected staining is observedwhich cannot be explained by variations in laboratory procedures and a problem with the product issuspected, contact Dako Technical Support.(125835-001)P04120EFG 01 GV800/2015.05 p. 8/32
Reagent PreparationA.1 EnVision FLEX Target Retrieval Solution, High pH (50x) (DM848, Code GV804) orEnVision FLEX Target Retrieval Solution, Low pH (50x) (DM849, Code GV805)Prepare a working solution by diluting the EnVision FLEX Target Retrieval Solution, High pH (50x) orLow pH (50x) concentrate 1:50 in distilled or deionized water. The following procedure isrecommended:1. Fill a Dako Omnis bulk bottle to the fill line with distilled or deionized water (3.325 L). Ensure toplace the bulk bottle on a horizontal surface before filling.2. Empty one 68 mL EnVision FLEX Target Retrieval Solution, High pH (50x) or Low pH (50x) (DakoOmnis) concentrate bottle into the bulk bottle.3. Transfer the detachable label from the concentrate bottle and attach it to the bulk bottle.4. Fasten the lid and gently invert the bulk bottle 2-3 times.Unused diluted solution may be stored at 2-8 C for one month. The diluted solution can be usedwithin 7 days when stored onboard the Dako Omnis instrument. Discard solution if cloudy inappearance.A.2 Wash Buffer (20x) (DM851, Code GC807)Prepare a working solution by diluting the Wash Buffer (20x) (Dako Omnis) concentrate 1:20 in DakoOmnis bulk bottle using distilled or deionized water. To minimize foaming the following procedure isrecommended:1. Fill a Dako Omnis bulk bottle to the fill line with distilled or deionized water (3.325 L). Ensure toplace the bulk bottle on a horizontal surface before filling.2. Empty one 175 mL Wash Buffer (20x) (Dako Omnis) concentrate bottle into the bulk bottle.3. Transfer the detachable label from the concentrate bottle and attach it to the bulk bottle.4. Fasten the lid and gently invert the bulk bottle 2-3 times.Unused diluted solution may be stored at 2-8 C for one month. The diluted solution can be usedwithin 7 days when stored onboard the Dako Omnis instrument. Remaining onboard stability is trackedby the Dako Omnis software. Discard solution if cloudy in appearance.A.3 EnVision FLEX Substrate Working SolutionThe DAB-containing EnVision FLEX Substrate Working Solution is prepared onboard the Dako Omnisinstrument. The automated process mixes one part of EnVision FLEX DAB Chromogen (DM847) with50 parts of EnVision FLE
EnVision FLEX DAB Chromogen and EnVision FLEX Su bstrate Buffer is 75 hours. Onboard stability of diluted EnVision FLEX Target Retrieva l Solution, High pH and Low pH is 7 days. Remaining onboard stability is tracked by the Dako Omnis software.
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