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Dako envision flex detection system

Manufactured by Agilent Technologies
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Sourced in Denmark

The Dako EnVision FLEX detection system is a versatile tool used in immunohistochemistry (IHC) and in situ hybridization (ISH) applications. It provides a sensitive method for the visualization of target molecules in biological samples.

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Lab products found in correlation

Dako omnis, by Agilent Technologies (2 mentions) Dako autostainer plus, by Agilent Technologies (2 mentions) Clone do 7, by Agilent Technologies (1 mentions) Clone ve1, by Abcam (1 mentions) Background sniper, by Biocare Medical (1 mentions) Dab chromogen kit, by Biocare Medical (1 mentions) Powervision detection system, by Immunologic (1 mentions) Ti e microscope, by Nikon (1 mentions) Nis elements basic research software, by Nikon (1 mentions) Cytoseal xyl, by Thermo Fisher Scientific (1 mentions) Target retrieval solution, by Agilent Technologies (1 mentions) Autostainer, by Agilent Technologies (1 mentions) Pt link pre treatment module, by Agilent Technologies (1 mentions) Meyer s haematoxylin, by Agilent Technologies (1 mentions)

Close spelling variants of this product

Envision System (550 citations) Envision Detection System (201 citations) EnVision FLEX (142 citations) Dako Envision system (132 citations) EnVision FLEX system (59 citations) EnVision FLEX detection system (24 citations) DAKO EnVision Detection System (22 citations) Dako EnVision Flex (10 citations) Dako EnVision Flex + System (7 citations)

8 protocols using dako envision flex detection system

1

Immunohistochemical Analysis of LIGHT

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BM biopsies were fixed in 4% neutral buffered formalin solution, decalcified, paraffin-embedded, and cut into 5 μm slices. After standard antigen retrieval procedures, the sections were incubated overnight at 4°C with primary mouse anti-human LIGHT (Abcam, Cambridge Science Park). The reaction was revealed with Dako EnVisionTM FLEX+ detection system (Dako Italia S.p.A. Milan, Italy).
Brunetti G., Rizzi R., Oranger A., Gigante I., Mori G., Taurino G., Mongelli T., Colaianni G., Di Benedetto A., Tamma R., Ingravallo G., Napoli A., Faienza M.F., Mestice A., Curci P., Specchia G., Colucci S, & Grano M. (2014). LIGHT/TNFSF14 increases osteoclastogenesis and decreases osteoblastogenesis in multiple myeloma-bone disease. Oncotarget, 5(24), 12950-12967.
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2

EGFR Assessment by IHC in Tissue

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Methods for EGFR reverse transcription polymerase chain reaction and amplification by fluorescence in situ hybridization13 (link) were previously published. An IHC assay was performed on FFPE tissue to assess EGFR protein expression using the EGFR E30 clone (Agilent, Santa Clara, CA) and the Dako EnVisionTM FLEX detection system on the Dako Link 48 automated staining platform (Agilent). The E30 clone recognizes both wild-type and EGFRvIII forms of EGFR. A trained pathologist evaluated EGFR IHC staining by scoring the percentage of positive cells in neoplastic cells at each intensity, which is categorized into 0 intensity (negative), 1+ intensity (weak), 2+ intensity (moderate), and 3+ intensity (strong).
Carneiro B.A., Papadopoulos K.P., Strickler J.H., Lassman A.B., Waqar S.N., Chae Y.K., Patel J.D., Shacham-Shmueli E., Kelly K., Khasraw M., Bestvina C.M., Merrell R., Huang K., Atluri H., Ansell P., Li R., Jin J., Anderson M.G., Reilly E.B., Morrison-Thiele G., Patel K., Robinson R.R., Aristide M.R, & Gan H.K. (2022). Phase I study of anti-epidermal growth factor receptor antibody-drug conjugate serclutamab talirine: Safety, pharmacokinetics, and antitumor activity in advanced glioblastoma. Neuro-Oncology Advances, 5(1), vdac183.
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3

BRAF and p53 IHC Staining in FFPE

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Sections (4 µm thick) of the original FFPE block used for molecular analysis were subjected to anti-BRAF (mutated V600E) IHC staining (cat. no. ab228461; clone VE1; dilution, 1/100; Abcam) and anti-p53 IHC staining (cat. no. M7001; clone DO-7; dilution, 1/200; Agilent Technologies, Inc.) on a Dako Omnis (Agilent Technologies, Inc.). Heat-induced epitope retrieval was performed using Dako Target Retrieval Solution pH 9 (cat. no. GV804; Agilent Technologies, Inc.) 30 min at 97˚C, followed by primary antibody incubation 20 min at 32˚C and detection with Dako Envision Flex detection system (cat. no. GV800; Agilent Technologies, Inc.) according to the manufacturer's protocol. The sections were counterstained with hematoxylin (cat. no. GC808; Agilent Technologies, Inc.). Control tissues (tonsil for p53 IHC and BRAF V600E mutated tumor for BRAF IHC) and universal negative control antibody (negative control Mouse IgG1; cat. no. X0931; dilution, 1/200; Agilent Technologies, Inc.) were processed in parallel with tissues exposed to the primary as described above.
Rusu S., Verocq C., Trepant A.L., Maris C., De Nève N., Blanchard O., Van Campenhout C., De Clercq S., Rorive S., Cotoi O.S., Decaestecker C., Salmon I, & D'Haene N. (2021). Immunohistochemistry as an accurate tool for the assessment of BRAF V600E and TP53 mutations in primary and metastatic melanoma. Molecular and Clinical Oncology, 15(6), 270.
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4

Immunohistochemical Analysis of TRK Receptors

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IHC for TRKA, TRKB, and TRKC expression was performed with a pan-TRK monoclonal antibody (mAb) (clone EPR17341; ref. ab181560 (Abcam, Cambridge, UK)). The antibody is reactive to a homologous region of TRKA, TRKB, and TRKC near the C-terminus. Appendix was used as positive control. All assays were performed on a Dako Omnis automated stainer platform (Agilent Technologies Inc.). Prior to staining, paraffin-embedded tissue sections were subjected to deparaffinization and hydration followed by heat-induced epitope retrieval in Dako Target Retrieval Solution pH9 (Agilent Technologies, Inc., ref. GV804) 30 min at 97°C. Pan-TRK antibody was diluted 1/200 and incubated for 20 min at 32°C. Detection was performed with Dako Envision Flex detection system (Agilent Technologies, Inc., ref. GV800) according to the manufacturer’s protocol. The sections were counterstained with hematoxylin (Agilent Technologies, Inc., ref. GC808).
Staining intensity (negative, weak, moderate, or strong), pattern (diffuse, focal, or rare positive cells), and localization (cytoplasmic or nuclear) were evaluated.
Demols A., Rocq L., Perez-Casanova L., Charry M., De Nève N., Ramadhan A., Van Campenhout C., De Clercq S., Maris C., Closset J., Lucidi V., Salmon I, & D’Haene N. (2023). A Two-Step Diagnostic Approach for NTRK Gene Fusion Detection in Biliary Tract and Pancreatic Adenocarcinomas. The Oncologist, 28(7), e520-e525.
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5

Immunohistochemical Analysis of HCMV

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All stains (except for the CNS DLBCL-classification) were carried out manually. Briefly, unspecific binding sites were blocked with Background Sniper (Biocare Medical, Concord, CA) for 10 min, and sections were incubated with primary Ab overnight at 4°C. PowerVision detection system (Immunologic, Duiven, The Netherlands) was used with a Romulin AEC or DAB chromogen kit (Biocare Medical) for antigen detection. The newly cut sections of the included TMAs were stained with the selected HCMV Abs (Table 3). The newly cut sections of the CNS DLBCL block were stained using the Dako Autostainer Plus (DakoCytomation) with Abs selected for diagnostics (Table 3). Dako EnVision FLEX detection system (DakoCytomation) was used subsequently for the visualization of the staining results, according to the manufacturer's instructions.
Libard S., Popova S.N., Amini R.M., Kärjä V., Pietiläinen T., Hämäläinen K.M., Sundström C., Hesselager G., Bergqvist M., Ekman S., Zetterling M., Smits A., Nilsson P., Pfeifer S., de Ståhl T.D., Enblad G., Ponten F, & Alafuzoff I. (2014). Human Cytomegalovirus Tegument Protein pp65 Is Detected in All Intra- and Extra-Axial Brain Tumours Independent of the Tumour Type or Grade. PLoS ONE, 9(9), e108861.
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6

Immunohistochemical Analysis of GPR64 Expression

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The paraffin-embedded tissues were sectioned (5 μm thick) and immunostained using the Dako EnVision FLEX + Detection system (DAKO, Carpinteria, CA, USA; #K8000). Antigen retrieval was performed by heating at low pH for 20 min and sections were rinsed in Dako wash buffer according to the manufacturer’s instructions. Endogenous peroxidase activity was blocked with Peroxidase-Blocking Reagent (10 min) before incubation with rabbit anti-GPR64 antibody at 1:400 for 20 min at room temperature. The primary antibody signal was amplified by EnVision FLEX + Rabbit (LINKER) before incubation with EnVision FLEX/HRP Detection Reagent for 30 min. Finally, sections were stained with 3,3’-diaminobenzidine (DAB) followed by counterstaining with Dako FLEX hematoxylin. Slides were rinsed and mounted in Cytoseal XYL (Thermo Scientific, Waltham, MA, USA). H&E staining was performed as described.(20 (link)) The bright field imaging was conducted by 40 oil objective (1.4 NA) on a Nikon Ti-E microscope equipped with 16.2 MegaPixels DS-Ri2 camera. DAB-stained pixels were defined for the Nikon NIS-Elements Basic Research software and were used for calculating the stained area fraction by ROI statistics module, as depicted in Supporting Fig. 5.
Balenga N., Azimzadeh P., Hogue J.A., Staats P.N., Shi Y., Koh J., Dressman H, & Olson JA J.r. (2016). Orphan Adhesion GPCR GPR64/ADGRG2 Is Overexpressed in Parathyroid Tumors and Attenuates Calcium-Sensing Receptor-Mediated Signaling. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 32(3), 654-666.
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7

Immunohistochemical Evaluation of Liver Markers

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Immunohistochemical reactions were performed on 4-μm formalin-fixed and paraffin-embedded tissue sections. Sections were stained on a Dako Autostainer with the Dako EnVision™ FLEX+ detection system (Dako, Glostrup, Denmark). The system detects primary mouse and rabbit antibodies and the reaction is visualised by EnVision™ FLEX DAB+ Chromogen. Using EnVision™ FLEX+ Mouse (LINKER) or EnVision™ FLEX+ Rabbit (LINKER) (Code K8019) signal amplification of primary mouse antibodies or primary rabbit antibodies were presented, respectively.
The deparaffinization, rehydration, and heat-induced epitope-retrieval (HIER) were carried out in one step with the 3-in-1 procedure buffer (Dako Target Retrieval Solution), pH 9 high ((10x)(3-in-1) Code S2375)) or pH 6 low, [(10x)(3-in-1) Code S1699)] at 97°C using a PT Link, Pre-Treatment Module (Dako). Tissue samples were analysed by light microscopy after 8 minutes counterstaining with Meyer’s haematoxylin (Dako). As primary antibodies we used Anti-CK19, −Glypican-3 and -AFP (Table 1).

Antibodies used in this study

AntibodyClonalityBufferDilutionIncubation
Anti-CK19monoclonal mousehighReady to use20 min(IR 615 Dako)
Anti-AFPpolyclonal rabbithighReady to use30 min(IR 6500 Dako)
Anti-Glypican-3monoclonal mouselowReady to use30 min(G1829R06 DCS Immunoline)
Bremmer F., Ströbel P., Jarry H., Strecker J., Gaisa N., Strauß A., Schweyer S., Radzun H.J, & Behnes C.L. (2015). CK19 is a sensitive marker for yolk sac tumours of the testis. Diagnostic Pathology, 10, 7.
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8

Optimized IHC Staining for Alzheimer's Aβ

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The IHC stainings were performed using automatic platform. The antibodies used and the pre-treatments applied are summarized in Table 1. Two of the antibodies, the pAβ1E4E11 and the Aβ7H3D6, a variant that recognize Aβ with unmodified N-terminus (umAβ), were both generated as previously described [32 (link)]. For these two antibodies, additional treatments were implemented following systematic testing in order to reach optimal results. The stainings were performed using Dako Autostainer Plus (DakoCytomation, Glostrup, Denmark) with the Dako EnVision Flex detection system (DakoCytomation), according to the manufacturer’s instructions.
Libard S., Walter J, & Alafuzoff I. (2021). In vivo Characterization of Biochemical Variants of Amyloid-β in Subjects with Idiopathic Normal Pressure Hydrocephalus and Alzheimer’s Disease Neuropathological Change. Journal of Alzheimer's Disease, 80(3), 1003-1012.
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