Dako omnis platform

Manufactured by Agilent Technologies

Sourced in United States, Denmark

The Dako Omnis platform is a comprehensive, automated immunohistochemistry (IHC) and in situ hybridization (ISH) solution for clinical laboratory applications. The platform is designed to streamline the staining process, ensuring consistent and reliable results.

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Lab products found in correlation

22c3 pharmdx antibody, by Agilent Technologies (3 mentions) 28 8 pharmdx antibody, by Agilent Technologies (2 mentions) Autostainer link 48, by Agilent Technologies (1 mentions) Mib 1, by Agilent Technologies (1 mentions) Dako envision flex, by Agilent Technologies (1 mentions) Anti cd8 antibody clone c8 144b, by Agilent Technologies (1 mentions) Envision flex, by Agilent Technologies (1 mentions) Aperio cs2, by Leica (1 mentions)

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Dako Omnis (50 citations) Omnis platform (15 citations) Dako Omnis automatic platform (2 citations)

15 protocols using dako omnis platform

Oncopanel Protein Expression Analysis

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The oncopanel in this study assessed the surface expression of 11 proteins (EGFR, MET, HER2, ALK, VEGFR, PDGFR, FLI1, KIT (CD117), CD30, AR, PD-L1) as previously published (15). The staining was performed on the Dako Omnis platform (Dako, Glostrup, Denmark) as previously published [14 (link),15 (link)]. Due to the very low prevalence of BRAF V600E in non-melanocytic head and neck carcinoma, we modified the oncopanel and excluded this marker. We also excluded PD-1 as a marker because PD-L1 expression has been established as a predictor of response to anti-PD-1 therapy and made the routine assessment of PD-1 expression redundant [6 (link)]. Briefly, we used formalin-fixed paraffin-embedded tissues and antibody details are provided in the Supplementary Table S1. Cases were considered positive if more than 10% of tumor cells showed immunopositivity in the appropriate cellular compartment. PD-L1 expression was scored according to the combined positive score (CPS) and cases with a CPS ≥ 1 were considered positive.

von Witzleben A., Müller-Richter U., Maurus K., Brändlein S., Theodoraki M.N., Brunner C., Laban S., Lennerz J., Möller P., Hoffmann T.K., Doescher J, & Schuler P.J. (2022). Protein-Based Oncopanel as Addition to Target Sequencing in Head and Neck Squamous Cell Carcinoma to Individualize Treatment Decisions. International Journal of Molecular Sciences, 23(24), 15835.

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Immunohistochemical Analysis of PD-L1 and CD20 in FFPE Tissue

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Paraffin-embedded tissue sections of patients fixed in formalin were cut into 4µm, the slides were processed and stained by the indirect immunoperoxidase method using the Dako Omnis Platform (Dako, Santa Clara, CA, USA). The slides underwent deparaffinization, followed by antigen retrieval with EnVision FLEX Target Retrieval Solution, Low pH for PD-L1, and High pH for CD20. The sections were incubated with primary antibodies Monoclonal Mouse Anti-Human PD-L1 (Clone 22C3; Dako Omnis; diluited at 1:30) and Monoclonal Mouse Anti-Human CD20cy (Clone L26; Dako Omnis; ready-to-Use). After endogenous peroxidase activity blocking with EnVision FLEX Peroxidase-Blocking Reagent, the products of the antigen–antibody reactions were visualized by EnVision FLEX DAB + Chromogen and EnVsion FLEX Substrate Buffer. Cell nuclei were stained with Hematoxylin (8 min). Tonsil FFPE sections were used as positive controls. For negative control, the primary antibody was omitted.

Rosamaria P., Daniela P., Rosanna L., Michele M., Annamaria C., Pamela P., Antonietta B.M., Alfredo Z.F., Gabriella D.B., Antonia Z., Stefania T, & Simona D.S. (2019). KRAS-Driven Lung Adenocarcinoma and B Cell Infiltration: Novel Insights for Immunotherapy. Cancers, 11(8), 1145.

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Oncopanel Protein Expression Analysis

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The oncopanel assessed expression of ten proteins from May 2014 to July 2016 and has been supplemented with PD-1 and PD-L1 since August 2016 (= 12 markers). In total, 45 patients were analyzed, 24 of them with the 10-marker panel and another 21 with the 12-marker panel. Staining was performed on the Dako Omnis platform (Dako, Glostrup, Denmark) as previously published [7] . Briefly, we used formalin-fixed paraffin-embedded tissues and antibody details are provided in Supplementary Table 1. Cases were considered positive if more than 10% of tumor cells showed immunopositivity in the appropriate cellular compartment (Supplementary Table 1). Representative histological images are displayed in Figure 1. Repeat oncopanel testing was performed in four patients (Table 2).

Doescher J., Weissinger S.E., Schönsteiner S.S., Lisson C., Bullinger L., Barth T.F., Leithäuser F., Mueller-Richter U., Laban S., Hoffmann T.K., Möller P., Lennerz J.K, & Schuler P.J. (2019). Clinical utility of a protein-based oncopanel in patients with end-stage head and neck cancer. Immunotherapy, 11(14).

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PD-L1 Expression Quantification Protocol

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The expression of PD-L1 on the surface of tumor cells was reported as a standard of care before treatment, which was performed using the 22C3 PharmDx antibody on the Dako Omnis platform (Agilent) and scored by published guidelines (26 ).

Abdelfatah E., Long M.D., Kajihara R., Oba T., Yamauchi T., Chen H., Sarkar J., Attwood K., Matsuzaki J., Segal B.H., Dy G.K, & Ito F. (2023). Predictive and Prognostic Implications of Circulating CX3CR1+ CD8+ T Cells in Non–Small Cell Lung Cancer Patients Treated with Chemo-Immunotherapy. Cancer Research Communications, 3(3), 510-520.

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BCL-2 Immunohistochemistry in Bone Marrow Biopsies

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Bone marrow core biopsies obtained at diagnosis were collected and analyzed for BCL-2 protein expression by immunohistochemistry (IHC), which was performed on a Dako Omnis platform (Agilent Technologies, Santa Clara, CA, USA) using the corresponding anti-BCL-2 antibody (clone 124). The percentage of BCL-2-expressing blast cells was manually scored under a light microscope by two independent observers (I.D.H. and A.D.) using the H-method, which is a simple, validated semi-quantitative immunostaining score of intensity and extent [22 (link)]. Intensity was graded on a scale of 0 to 3 (0 = absent, 1 = weak, 2 = moderate, and 3 = intense). Extent of staining was scored from 0 to 100, which refers to the percentage of blast cells staining positive for BCL-2. Intensity and extent scores were multiplied to generate an H-score ranging from 0 to 300. Based on previous research [23 (link)], we established an H-score of 20 as cut-off to discriminate between BCL-2⁻ and BCL-2⁺ samples.

haes I.D., Dendooven A., Mercier M.L., Puylaert P., Vermeulen K., Kockx M., Deiteren K., Maes M.B., Berneman Z, & Anguille S. (2020). Absence of BCL-2 Expression Identifies a Subgroup of AML with Distinct Phenotypic, Molecular, and Clinical Characteristics. Journal of Clinical Medicine, 9(10), 3090.

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PD-L1 Immunohistochemistry Diagnostic Protocols

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In melanoma samples, PD-L1 expression was assessed using the Dako Omnis platform (Agilent, Santa Clara, CA) and the 28–8 pharmDx antibody (Agilent, Santa Clara, CA), which is the FDA-approved complementary diagnostic for nivolumab. For RCC and NSCLC samples, the 22c3 pharmDx antibody (Agilent, Santa Clara, CA) was employed on Autostainer Link 48 (Agilent, Santa Clara, CA), which is the FDA-approved companion diagnostic for pembrolizumab. Established cutoffs for the diagnostics in each histologic type were used to score PD-L1 IHC tumor proportion score (TPS) and immune cell staining (ICS) as follows: melanoma TPS, 1%, [13 ] NSCLC TPS, 50 and 1%, [7 ] RCC TPS, 1%; RCC ICS, 1%.

Conroy J.M., Pabla S., Nesline M.K., Glenn S.T., Papanicolau-Sengos A., Burgher B., Andreas J., Giamo V., Wang Y., Lenzo F.L., Bshara W., Khalil M., Dy G.K., Madden K.G., Shirai K., Dragnev K., Tafe L.J., Zhu J., Labriola M., Marin D., McCall S.J., Clarke J., George D.J., Zhang T., Zibelman M., Ghatalia P., Araujo-Fernandez I., de la Cruz-Merino L., Singavi A., George B., MacKinnon A.C., Thompson J., Singh R., Jacob R., Kasuganti D., Shah N., Day R., Galluzzi L., Gardner M, & Morrison C. (2019). Next generation sequencing of PD-L1 for predicting response to immune checkpoint inhibitors. Journal for Immunotherapy of Cancer, 7, 18.

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PD-L1 and CD8+ T-cell Tumor Infiltration

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The expression of PD-L1 on the surface of tumor cells was assessed in all samples via the Dako Omnis platform (Agilent, Santa Clara, CA) using the anti-PD-L1 22C3 pharmDx antibody (Agilent, Santa Clara, CA), as per manufacturers protocol (22 ). Expression levels were scored as per published guidelines (23 ). Additional serially sectioned tissue was evaluated for lymphocyte infiltration using the anti-CD8 antibody C8/144B (Agilent, Santa Clara, CA) at a dilution ratio of 1:75 and assigned a qualitative score of non-infiltrated, infiltrated, or excluded. Non-infiltrated referred to a sparse number of CD8⁺ T-cells that infiltrate nests of neoplastic cells in a non-overlapping fashion and with less than 5% of the tumor showing an infiltrating pattern. Infiltrated represents frequent CD8⁺ T-cells that infiltrate nests of neoplastic cells in an overlapping fashion at least focally and in more than 5% of the tumor. Excluded represents restriction of more than 95% of all CD8⁺ T-cells in a tumor to the periphery or interstitial stromal areas and not actively invading nest or groups of neoplastic cells.

Lenzo F.L., Kato S., Pabla S., DePietro P., Nesline M.K., Conroy J.M., Burgher B., Glenn S.T., Kuvshinoff B., Kurzrock R, & Morrison C. (2021). Immune profiling and immunotherapeutic targets in pancreatic cancer. Annals of Translational Medicine, 9(2), 119.

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PD-L1 Immunohistochemical Detection

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Immunohistochemical (IHC) detection of tumour PD-L1 expression was performed using the PD-L1 Clone 22C3 kit (pharmDx, Agilent Technologies, Santa Clara, CA, USA) and the Dako Omnis platform (Agilent Technologies, Carpinteria, CA, USA). The percentage of tumour cells with PD-L1 expression (positive membrane staining) was obtained by counting at least 100 viable cells, and this was the so-called TPS. The evaluation of PD-L1 expression followed the specific requests of the treating clinician in terms of selection of the tested cohort and timing (diagnostic biopsy or rebiopsy).

Bironzo P., Reale M.L., Sperone T., Tabbò F., Caglio A., Listì A., Passiglia F., Di Maio M., Righi L., Bussolino F., Scagliotti G.V, & Novello S. (2021). Clinical and Molecular Features of Epidermal Growth Factor Receptor (EGFR) Mutation Positive Non-Small-Cell Lung Cancer (NSCLC) Patients Treated with Tyrosine Kinase Inhibitors (TKIs): Predictive and Prognostic Role of Co-Mutations. Cancers, 13(10), 2425.

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Evaluating PD-L1 and CD8+ T-cell Infiltration

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The expression of PD-L1 on the surface of tumor cells was assessed in all samples by means of the Dako Omnis platform (Agilent, Santa Clara, CA) and the 28–8 pharmDx antibody. Expression levels, and were scored as per published guidelines [19 ]. Additional serially sectioned tissue was evaluated for lymphocyte infiltration using the anti-CD8 antibody C8/144B (Agilent, Santa Clara, CA) and assigned a qualitative score of non-infiltrated, infiltrated, or excluded. Non-infiltrated referred to a sparse number of CD8⁺ T-cells that infiltrate nests of neoplastic cells and with less than 5% of the tumor showing an infiltrating pattern. Infiltrated represents frequent CD8⁺ T-cells that infiltrate nests of neoplastic cells in an overlapping fashion at least focally and in more than 5% of the tumor. Excluded represents restriction of more than 95% of all CD8⁺ T-cells in a tumor to the periphery or interstitial stromal areas and not actively invading nest or groups of neoplastic cells.

Morrison C., Pabla S., Conroy J.M., Nesline M.K., Glenn S.T., Dressman D., Papanicolau-Sengos A., Burgher B., Andreas J., Giamo V., Qin M., Wang Y., Lenzo F.L., Omilian A., Bshara W., Zibelman M., Ghatalia P., Dragnev K., Shirai K., Madden K.G., Tafe L.J., Shah N., Kasuganti D., de la Cruz-Merino L., Araujo I., Saenger Y., Bogardus M., Villalona-Calero M., Diaz Z., Day R., Eisenberg M., Anderson S.M., Puzanov I., Galluzzi L., Gardner M, & Ernstoff M.S. (2018). Predicting response to checkpoint inhibitors in melanoma beyond PD-L1 and mutational burden. Journal for Immunotherapy of Cancer, 6, 32.

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Immunohistochemical Analysis of MCM3 and Ki-67

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IHC was performed on tissue microarray sections of 4 μm thickness using a DAKO Omnis platform (Agilent Technologies, Santa Clara, CA). Heat-induced epitope retrieval was performed on board, followed by antibody incubation in Dako EnVision FLEX (Agilent Technologies, Santa Clara, CA). For MCM3, the DAKO Omnis protocol H30-X-30 was utilized with recombinant rabbit monoclonal anti-MCM3 antibody (dilution 1/1000, clone EPR7080, catalogue # ab128923; Abcam, Cambridge, UK). For Ki-67, the DAKO Omnis protocol L20-X-20 was used with mouse monoclonal anti-Ki-67 antibody (ready-to-use, clone Mib-1, catalogue # GA626; Agilent Technologies, Santa Clara, CA). Nuclear expression in tumor cells was scored in 5% increments. Based on the distribution, no naturally occurring cut-off was apparent. Therefore, scores were categorized into 3 relatively equally sized groups after several iterations to optimize for prognostic stratification (For MCM3: <40%, 40% to 75%, >75%; for Ki-67 <20%, 20% to 30%, >30%). TMA cores with less than 10% tumor content were excluded from study. A subset of cases was scored again by a second observer blinded to the initial scores. Previously generated immunohistochemical and chromogenic in situ hybridization (CISH) data for p53, p16, RB1, and CCNE1 were used for correlative analysis [5 (link), 18 (link), 21 (link)].

Kang E.Y., Millstein J., Popovic G., Meagher N.S., Bolithon A., Talhouk A., Chiu D.S., Anglesio M.S., Leung B., Tang K., Lambie N., Pavanello M., Da-anoy A., Lambrechts D., Loverix L., Olbrecht S., Bisinotto C., Garcia-Donas J., Ruiz-Llorente S., Yagüe-Fernandez M., Edwards R.P., Elishaev E., Olawaiye A., Taylor S., Ataseven B., du Bois A., Harter P., Lester J., Høgdall C.K., Armasu S.M., Huang Y., Vierkant R.A., Wang C., Winham S.J., Heublein S., Kommoss F.K., Cramer D.W., Sasamoto N., van-Wagensveld L., Lycke M., Mateoiu C., Joseph J., Pike M.C., Odunsi K., Tseng C.C., Pearce C.L., Bilic S., Conrads T.P., Hartmann A., Hein A., Jones M.E., Leung Y., Beckmann M.W., Ruebner M., Schoemaker M.J., Terry K.L., El-Bahrawy M.A., Coulson P., Etter J.L., LaVigne-Mager K., Andress J., Grube M., Fischer A., Neudeck N., Robertson G., Farrell R., Barlow E., Quinn C., Hettiaratchi A., Casablanca Y., Erber R., Stewart C.J., Tan A., Yu Y., Boros J., Brand A.H., Harnett P.R., Kennedy C.J., Nevins N., Morgan T., Fasching P.A., Vergote I., Swerdlow A.J., dos Reis F.J., Maxwell G.L., Neuhausen S.L., Barquin-Garcia A., Modugno F., Moysich K.B., Crowe P.J., Hirasawa A., Heitz F., Karlan B.Y., Goode E.L., Sinn P., Horlings H.M., Høgdall E., Sundfeldt K., Kommoss S., Staebler A., Wu A.H., Cohen P.A., DeFazio A., Lee C.H., Steed H., Le N.D., Gayther S.A., Lawrenson K., Pharoah P.D., Konecny G., Cook L.S., Ramus S.J., Kelemen L.E, & Köbel M. (2021). MCM3 is a novel proliferation marker associated with longer survival for patients with tubo-ovarian high-grade serous carcinoma. Virchows Archiv : an international journal of pathology, 480(4), 855-871.

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