Optiview dab detection kit

Manufactured by Roche

Sourced in United States, Switzerland

The OptiView DAB Detection Kit is a reagent system used in immunohistochemistry (IHC) procedures. It is designed to detect and visualize target proteins in tissue samples. The kit provides a series of reagents that enable the localization and identification of specific antigens through a chromogenic detection process. The core function of the OptiView DAB Detection Kit is to facilitate the visualization of target analytes in prepared tissue samples.

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Lab products found in correlation

Benchmark ultra, by Roche (17 mentions) Ez prep solution, by Roche (13 mentions) Hematoxylin 2, by Roche (12 mentions) Ventana benchmark xt, by Roche (10 mentions) Hematoxylin and bluing reagent, by Roche (9 mentions) Cell conditioning 1 cc1, by Roche (8 mentions) Benchmark xt, by Roche (7 mentions) Benchmark ultra platform, by Roche (6 mentions) Benchmark ultra autostainer, by Roche (4 mentions) Positive charged slides, by Bio-Optica (4 mentions) Benchmark gx, by Roche (4 mentions) Ventana benchmark ultra, by Roche (3 mentions) Cell condition 1 solution, by Roche (3 mentions) Ez prep, by Roche (3 mentions) Pd l1 clone 22c3, by Agilent Technologies (3 mentions) Clone 22c3, by Agilent Technologies (3 mentions) Cc1 solution, by Roche (3 mentions) Optiview amplification kit, by Roche (3 mentions) Anti nanog antibody, by Cell Signaling Technology (2 mentions) Protease 1, by Roche (2 mentions)

87 protocols using optiview dab detection kit

Immunohistochemical Analysis of NANOG

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For immunohistochemistry, 3-4 μm thick slides were cut from FFPE tissue blocks, followed by automated antigen retrieval (BenchMark ULTRA, Ventana, Tuscon, AZ, USA) and staining with commercially available anti-NANOG antibodies (Cell Signaling, cat no. 4903, dilution 1 : 200) (Merck, Kenilworth, New Jersey, USA). Reactions were visualized by incubation with peroxidase and 3,3′-diaminobenzidine (OptiVIEW DAB Detection Kit, Roche, Basel, Switzerland) and then counterstained with hematoxylin. Negative controls omitting the primary antibody binding were included in every run of samples. Testicular seminoma served as a positive control.
We evaluated the extent of positive reaction semiquantitatively (negative-0%, below 25%, between 25 and 50%, between 50 and 75%, and above 75%), intensity (weak, moderate, and strong), and staining pattern (nuclear, cytoplasmic staining, or both).

Grubelnik G., Boštjančič E., Grošelj A, & Zidar N. (2020). Expression of NANOG and Its Regulation in Oral Squamous Cell Carcinoma. BioMed Research International, 2020, 8573793.

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Fused-USgFNAC Cytology and Immunohistochemistry for Head and Neck Tumors

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Cytological results in nodes for which Fused-USgFNAC was performed were refereed as reference standard. Part of the FNAC material was processed in smears, air-dried and stained with Giemsa stain. Another portion of every aspirate was fixed in 10 mL 4% formalin and embedded in paraffin for further immunohistochemistry examination if deemed necessary, according to routine diagnostic workup. All samples were evaluated by experienced head and neck pathologists in a clinical setting and the cytological results were used retrospectively. HPV status was assessed immunohistochemically on formalin-fixed paraffin-embedded tissue samples from tumor biopsies or resections during standard routine diagnostic procedures. Antibodies for p53 (DO-7, 1/7000, DAKO) and p16 (E6H4; ready to use, Ventana Medical systems/Roche/Arizona, USA) were used in a Benchmark ULTRA autostainer (Ventana Medical systems). Reactions were detected using the OptiView DAB Detection kit (#760-700; Roche) for visualization of p16 and p53. Finally, the slides were counterstained with Hematoxylin II and Bluing Reagent (Ventana Medical Systems).

de Koekkoek-Doll P.K., Roberti S., Smit L., Vogel W.V., Beets-Tan R., van den Brekel M.W, & Castelijns J. (2022). ADC Values of Cytologically Benign and Cytologically Malignant 18 F-FDG PET-Positive Lymph Nodes of Head and Neck Squamous Cell Carcinoma. Cancers, 14(16), 4019.

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Automated Immunohistochemistry for Androgen Receptor

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Immunohistochemical staining was performed by an automated slide staining instrument BenchMark ULTRA (Ventana Medical System/Roche, Tucson, Arizona, United States). For immunohistochemistry, slides were deparaffinized and rehydrated. AR antibody clone M AR 441 (Dako, Glostrup, Denmark) was used as primary antibody. For detection of the AR, tissues were pretreated by heat antigen retrieval with a Cell Conditioner 1 solution (Roche, Basel, Switzerland) for 64 min with protease 1 (Roche). AR antibodies were diluted 1:200 in an antibody-diluent and incubated for 32 min at 37 °C in the platform. For visualization, the indirect biotin-free OptiView DAB Detection Kit (Roche) was used. The slides were counterstained and mounted. Internal controls served as positive controls for AR.
In microscopic assessment, AR staining was distributed homogeneously. Expression was quantified according to a modified scoring system, which has already been used for assessment of AR immunoreaction. Diversity of positive cells was classified to a score 0–5 [23] (link) by a researcher blinded for diabetes status.

Lutz S.Z., Hennenlotter J., Scharpf M.O., Sailer C., Fritsche L., Schmid V., Kantartzis K., Wagner R., Lehmann R., Berti L., Peter A., Staiger H., Fritsche A., Fend F., Todenhöfer T., Stenzl A., Häring H.U, & Heni M. (2017). Androgen receptor overexpression in prostate cancer in type 2 diabetes. Molecular Metabolism, 8, 158-166.

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Fibulin-2 Immunohistochemistry in Tumors

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Whole tumour sections were used for fibulin-2 staining. Slides were placed on the Ventana Benchmark Ultra automated immuno-stainer (Roche) and dewaxed. Heat-induced epitope retrieval was performed with Ventana`s Ultra CC1 retrieval solution for 56 minutes at 99°C followed by incubation with a rabbit polyclonal anti-fibulin-2 antibody (Thermo Fisher, product number PA5-51665 and lot number A35146) at a 1:200 dilution in Ventana Antibody Diluent (251–018) for 32 minutes at 36°C. Presence of the antigen was visualized by using the OptiView DAB detection kit (Roche).

Klingen T.A., Chen Y., Aas H., Wik E, & Akslen L.A. (2021). Fibulin-2 expression associates with vascular invasion and patient survival in breast cancer. PLoS ONE, 16(4), e0249767.

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HER2 Immunohistochemistry and In Situ Hybridisation Scoring

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For HER2 IHC, the primary antibody was rabbit monoclonal RTU clone 4B5 (Ventana, Roche) with antigen retrieval: 32 min CC1, incubation time 24 min. and Optiview DAB detection kit, according to the manufacturer. HER2 IHC was scored positive when distinct membranous staining was observed in at least 10% of the tumour cells and was categorised by staining intensity (1+ to 3+). In addition, the H score (0–300) was calculated.
For HER2 in situ hybridisation (ISH), the dual colour HER2/Ch17 DISH or the mono colour HER2 SISH kit from Ventana were used according to instruction from the manufacturer. The mean number of HER2 copies per nucleus was calculated after counting 40 tumour cell nuclei. The copy number variation (CNV) was distributed in three categories: 1–4, 5–10, and >10. The ratio with Ch17 was not used because the Ch17 copy number could not be calculated for the complete set of samples.
All biopsies were centrally reviewed prior to study inclusion by one of two experienced thoracic pathologists (E.T. or K.M.).

de Langen A.J., Jebbink M., Hashemi S.M., Kuiper J.L., de Bruin-Visser J., Monkhorst K., Thunnissen E, & Smit E.F. (2018). Trastuzumab and paclitaxel in patients with EGFR mutated NSCLC that express HER2 after progression on EGFR TKI treatment. British Journal of Cancer, 119(5), 558-564.

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Histological Examination of Brain Tissue

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For histological examination of brains, tissue was fixed in 4% formaldehyde for at least 12 h. The tissue was dehydrated, embedded in paraffin, and sectioned at 2 μm according to standard protocols. Hematoxylin and eosin (HE) stainings were applied according to standard protocols. 3,3’-Diaminobenzidine (DAB) stainings were performed on a Ventana Benchmark system using the ultraView or OptiView DAB detection kit (all Roche Diagnostics, Basel, CH). The following antibodies were used: Cleaved Caspase-3 (CC-3): Cell Signaling #9664, RRID:AB_2070042 (1:100); GFP: Abcam #ab290, RRID:AB_303395 (1:500); Ki67: Abcam #ab15580, RRID:AB_443209 (1:100); MYC: Zeta Corporation #Z2734RL (1:25); Nestin: Abcam #ab221660, RRID:AB_2909415 (1:2000); NeuN: Merck #MAB377, RRID:AB_2298772 (1:50); OLIG2: Merck #AB9610, RRID:AB_570666 (1:200); SMARCA4: Abcam #ab110641, RRID:AB_10861578 (1:25); and SOX2: Abcam #92,494, RRID:AB_10585428 (1:200).

Göbel C., Godbole S., Schoof M., Holdhof D., Kresbach C., Loose C., Neumann J, & Schüller U. (2023). MYC overexpression and SMARCA4 loss cooperate to drive medulloblastoma formation in mice. Acta Neuropathologica Communications, 11, 174.

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Automated Immunohistochemical NANOG Evaluation

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Commercially available anti-NANOG antibody (Cell Signaling, cat no. 4903, dilution 1:200) (Merck, Kenilworth, New Jersey, USA) was used for immunohistochemistry. Automated antigen retrieval and staining (BenchMark ULTRA, Ventana, Tuscon, AZ, USA) were performed on unstained 4 µm thick slides cut from FFPE tissue blocks. Visualization of reaction was provided by peroxidase and 3,3′-diaminobenzidine incubation (OptiVIEW DAB Detection Kit, Roche, Basel, Switzerland) and after counterstaining with hematoxylin. Every run of the samples included a positive control (testicular seminoma) and negative controls omitting the primary antibody binding.
Using semi-quantitative approach, we evaluated the extent of staining (negative—score 0, below 25%—score 1, between 25 and 50%—score 2, between 50 and 75%—score 3 and above 75%—score 4). We also evaluated the intensity of staining (negative—score 0, weak—score 1, moderate—score 2, strong—score 3) and staining pattern (nuclear, cytoplasmic staining or both). We calculated the combined immunohistochemical score by multiplying the intensity and the extent scores.

Grubelnik G., Boštjančič E., Aničin A., Dovšak T, & Zidar N. (2021). MicroRNAs and Long Non-Coding RNAs as Regulators of NANOG Expression in the Development of Oral Squamous Cell Carcinoma. Frontiers in Oncology, 10, 579053.

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Automated IHC for SARS-CoV-2 Spike and Nucleocapsid

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Automated classical immunohistochemical (IHC) stains were performed using the Benchmark ULTRA (Roche, Ventana Medical Systems, Innovation Park Drive Tucson, Arizona 85755, USA) on formalin-fixed and paraffin-embedded (FFPE) tissue sections (3 μm). After dewaxing, tissue slides were heat pretreated using a CC1 (pH8) buffer (05424569001, Roche) for 64 min at 98 °C. The slides were blocked for endogenous peroxidase activity and incubated with primary anti–2019-nCOV Spike rabbit polyclonal (A20022, ABclonal, Diagomics, Blagnac, France) (1/100 in Envision FLEX antibody diluent, K800621-2, Agilent, Santa Clara, CA 95051, USA) and anti–SARS-CoV-2 nucleocapsid mouse monoclonal (Ready to Use, clone BSB-134, BioSB, Diagomics, Blagnac, France) antibodies for 64 and 92 min, respectively. Targets were then visualized using the OptiView DAB detection kit (06396500001, Roche). The tissue slides were counterstained using hematoxylin (05277965001, Roche) for 8 min followed by postcoloration using Bluing reagent for 4 min at room temperature (05266769001, Roche). The slides were then dehydrated (ethanol and xylene) and mounted using xylene-based mounting (TissueTek Prisma®, Sakura Finetek Europe B.V., The Netherlands).

Dubucs C., Groussolles M., Ousselin J., Sartor A., Van Acker N., Vayssière C., Pasquier C., Reyre J., Batlle L., Favarel clinical research associate S., Duchanois midwife D., Jauffret clinical research associate V., Courtade-Saïdi M, & Aziza J. (2022). Severe placental lesions due to maternal SARS-CoV-2 infection associated to intrauterine fetal death. Human Pathology, 121, 46-55.

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Immunohistochemical Staining for Cell Proliferation and Hepatocyte Markers

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Immunohistochemical staining of the serial sections was performed using an automated slide stainer (Benchmark Ultra; Ventana Medical Systems, Roche, Basel, Switzerland). All sections were deparaffinised and boiled in Ventana CC1 buffer (Ventana Medical Systems, Roche, Basel, Switzerland), pH 8, for heat-induced epitope retrieval. The sections were incubated for 32 min with two different antibodies. Ready-to-use monoclonal mouse anti-rat Ki-67 specific antibody (clone 30–9; Ventana Medical Systems, Roche, Basel, Switzerland) diluted at 1:20 was used as a proliferation marker, and anti-beta-catenin antibody (clone β-Catenin-1; Agilent Technologies, Santa Clara, CA, U.S.) diluted at 1:100 was used as a hepatocyte cell surface marker. The sections were counterstained with haematoxylin. An OptiView DAB Detection Kit (Roche, Basel, Switzerland) and UltraView Universal Alkaline Phosphatase Red Detection Kit (Roche, Basel, Switzerland) were then used for visualization of the bound antibodies.

Lund A., Andersen K.J., Meier M., Pedersen M.I., Knudsen A.R., Kirkegård J., Mortensen F.V, & Nyengaard J.R. (2023). Biochemical and morphological responses to post-hepatectomy liver failure in rats. Scientific Reports, 13, 13544.

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PD-L1 Expression Analysis in Bone Marrow

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PD‐L1 expression study was carried out using anti‐PD‐L1 antibody clone SP142 (Abcam, Cambridge, UK). Ten BM samples were also stained with antibody clone SP263 (Roche, Ventana, Tucson, AZ, USA) and the results were correlated with each other. In brief, BM sections of 5‐μm thickness were deparaffinized with EZ prep (Ventana Medical Systems, Inc., Tucson, AZ, USA) and underwent a 64‐min pretreatment using Cell Condition 1 solution (Ventana Medical Systems, Inc.). The slides were then incubated with anti‐PD‐L1 antibody for 120 min using the automated Ventana Benchmark XT. Labeling was detected with the Optiview DAB Detection Kit (Roche, Ventana, AZ, USA) as per the manufacturer's protocol. Dilutions were 1:100 for antibody SP142 and a ready‐to‐use kit for antibody SP263. All sections were counterstained with hematoxylin in Ventana reagent.

Lee S., Lin C., Wei C., Chang K., Yuan C., Tsai C., Liu J., Hou H., Tang J., Chou W, & Tien H. (2021). PD‐L1 expression in megakaryocytes and its clinicopathological features in primary myelofibrosis patients. The Journal of Pathology: Clinical Research, 8(1), 78-87.

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