Formalin-fixed Paraffin-embedded (FFPE) tissue sections orTMAs were cut at 4–5 µm on charged slides. Slides were dried at 65°C for 6 hours. After drying, the slides were placed on the BOND RXm Research Stainer (Leica Biosystems) and deparaffinized with BOND Dewax solution (AR9222, Lecia Biosystems). The multispectral immunofluorescent (mIF) staining process involved serial applications of the following for each biomarker: epitope retrieval/stripping with ER1 (citrate buffer pH 6, AR996, Leica Biosystems ), blocking buffer (AKOYA Biosciences), primary antibody, Opal Polymer HRP secondary antibody (AKOYA Biosciences), Opal Fluorophore (AKOYA Biosciences). All AKOYA reagents used for mIF staining come as a kit (NEL821001KT). Spectral DAPI (AKOYA Biosciences) was applied once slides were removed from the BOND. They were cover slipped using an aqueous method and Diamond antifade mounting medium (Invitrogen ThermoFisher). The mIF panel consisted of the following antibodies (clone, company, and Opal Fluorophores): Cyclin D (SP4, Epredia, Opal 570), Cyclin E (EP435E, abcam, Opal 520), MCM2 (RBT-MCM2, Biosb, Opal Polaris 480), Pan Cytokeratin (AE1AE3, Agilent DAKO, Opal Polaris 780), pHH3 (Ser10, Millipore Sigma, Opal 620), pRB (Ser807/811, Cell Signaling, Opal 690).
Opal polymer hrp secondary antibody
Manufactured by Akoya Biosciences
The Opal Polymer HRP secondary antibody is a laboratory reagent designed to be used in immunohistochemistry (IHC) and immunofluorescence (IF) applications. The product consists of a horseradish peroxidase (HRP) enzyme conjugated to a polymer backbone, which can bind to and amplify the signal from primary antibodies directed against target proteins or antigens of interest.
Automatically generated - may contain errors
Lab products found in correlation
Spectral dapi, by Akoya Biosciences (3 mentions)
Bond rxm research stainer, by Leica (3 mentions)
Diamond antifade mounting medium, by Thermo Fisher Scientific (3 mentions)
Opal fluorophore, by Akoya Biosciences (3 mentions)
Ae1 ae3, by Agilent Technologies (1 mentions)
Ccnd1, by Thermo Fisher Scientific (1 mentions)
Ar9640, by Leica (1 mentions)
3 protocols using opal polymer hrp secondary antibody
1
Multiplex Immunofluorescence for Cell Cycle Markers
2
Multiplex Immunofluorescence Staining of FFPE Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Formalin-fixed paraffin-embedded (FFPE) 4 µm sections were cut and placed on charged slides. Slides were dried at 65 °C for 2 h. After drying, the slides were placed on the BOND RXm Research Stainer (Leica Biosystems) and deparaffinized with BOND Dewax solution (AR9222, Lecia Biosystems). The multispectral immunofluorescent (mIF) staining process involved serial repetitions of the following for each biomarker: epitope retrieval/stripping with ER1 (citrate buffer pH 6, AR996, Leica Biosystems) or ER2 (Tris-EDTA buffer pH9, AR9640, Leica Biosystems), blocking buffer (AKOYA Biosciences), primary antibody, Opal Polymer HRP secondary antibody (AKOYA Biosciences), Opal Fluorophore (AKOYA Biosciences). All AKOYA reagents used for mIF staining come as a kit (NEL821001KT). Spectral DAPI (AKOYA Biosciences) was applied once slides were removed from the BOND. They were cover slipped using an aqueous method and Diamond antifade mounting medium (Invitrogen ThermoFisher). The mIF panel consisted of the following antibodies: Ki67 (Abcam, ab16667), AE1AE3 (Dako, M3515), CCNE (Abcam, ab33911), CCND1 (ThermoFisher, MA1-39546), CCNA (Abcam, ab32386), RB (Cell Signaling, 9309s), pRB (Cell Signaling, 8516), and MCM2 (BioSb, BSB6334).
3
Multiplex Immunofluorescence for Cell Cycle Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formalin-fixed Paraffin-embedded (FFPE) tissue sections orTMAs were cut at 4–5 µm on charged slides. Slides were dried at 65°C for 6 hours. After drying, the slides were placed on the BOND RXm Research Stainer (Leica Biosystems) and deparaffinized with BOND Dewax solution (AR9222, Lecia Biosystems). The multispectral immunofluorescent (mIF) staining process involved serial applications of the following for each biomarker: epitope retrieval/stripping with ER1 (citrate buffer pH 6, AR996, Leica Biosystems ), blocking buffer (AKOYA Biosciences), primary antibody, Opal Polymer HRP secondary antibody (AKOYA Biosciences), Opal Fluorophore (AKOYA Biosciences). All AKOYA reagents used for mIF staining come as a kit (NEL821001KT). Spectral DAPI (AKOYA Biosciences) was applied once slides were removed from the BOND. They were cover slipped using an aqueous method and Diamond antifade mounting medium (Invitrogen ThermoFisher). The mIF panel consisted of the following antibodies (clone, company, and Opal Fluorophores): Cyclin D (SP4, Epredia, Opal 570), Cyclin E (EP435E, abcam, Opal 520), MCM2 (RBT-MCM2, Biosb, Opal Polaris 480), Pan Cytokeratin (AE1AE3, Agilent DAKO, Opal Polaris 780), pHH3 (Ser10, Millipore Sigma, Opal 620), pRB (Ser807/811, Cell Signaling, Opal 690).
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