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Opal multiplex kit

Manufactured by PerkinElmer
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The OpalTM Multiplex Kit is a laboratory tool designed for the simultaneous detection and analysis of multiple protein targets in a single sample. It provides a standardized and reproducible method for performing multiplexed immunofluorescence analysis.

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Lab products found in correlation

Prolong diamond antifade mountant containing dapi, by Thermo Fisher Scientific (4 mentions) Anti cd19, by Biocare Medical (2 mentions) Anti bcl6, by Biocare Medical (2 mentions) Anti igd, by Agilent Technologies (2 mentions) Anti cd27, by Abcam (2 mentions) Vectra polaris, by PerkinElmer (2 mentions) Ab133616, by Abcam (1 mentions) Ab27988, by Abcam (1 mentions) Ab8021, by Abcam (1 mentions) Ab150440, by Abcam (1 mentions) Ab85792, by Abcam (1 mentions) Ab20181, by Abcam (1 mentions) Ab27969, by Abcam (1 mentions) Anti cd31, by Cell Signaling Technology (1 mentions) Hla dr, by Abcam (1 mentions) Pan ck, by Abcam (1 mentions) α sma, by Cell Signaling Technology (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) Vimentin, by Cell Signaling Technology (1 mentions) Anti cd3, by Agilent Technologies (1 mentions)

Close spelling variants of this product

Opal Multiplex IHC Kit (5 citations) Opal Multiplex fIHC kit (4 citations) Opal multiplex 6-plex kits (3 citations) Opal Multiplex tissue staining kit (3 citations) Opal Multiplex Immunohistochemistry Kits (2 citations) Opal Multiplex IHC assay kit (2 citations)

8 protocols using opal multiplex kit

1

Multiplex Immunofluorescence Profiling of Tissue Samples

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Tissue samples were fixed in formalin, embedded in paraffin, and sectioned. These specimens were incubated with the following antibodies: anti-CD3 (A045229–2; DAKO), anti-CD4 (ab133616; Abcam), anti-Pan-CK (ab27988; Abcam), anti-CD31 (3528; Cell Signaling Technology), anti-SLAMF7 (HPA055945; Sigma-Aldrich), anti-CX3CR1 (ab8021; Abcam), anti-T-bet (ab150440; Abcam), anti-GATA3 (MA1028; Invitrogen), anti-Ror gamma (ab212496; Abcam), anti-CXCR5 (clone: MAB190; R&D Systems), anti-FoxP3 (clone: 98377; Cell Signaling Technology), anti-CD8 (ab85792; Abcam), anti-PD1 (B13300; Lifespan Bioscience), anti-GZMB (ab4095; Abcam), anti-HLA-DR (ab20181; Abcam), anti-TGF-β (ab27969; Abcam) and anti-cleaved caspase-3 (9664; Cell Signaling Technology) followed by incubation with a secondary antibody using an OpalTM Multiplex Kit (Perkin Elmer). The samples were mounted with ProLongTM Diamond Antifade mountant containing DAPI (Invitrogen).
Kaneko N., Boucau J., Kuo H.H., Perugino C., Mahajan V.S., Farmer J.R., Liu H., Diefenbach T.J., Piechocka-Trocha A., Lefteri K., Waring M.T., Premo K.R., Walker B.D., Li J.Z., Gaiha G., Yu X.G., Lichterfeld M., Padera RF J.r, & Pillai S. (2022). Temporal changes in T cell subsets and expansion of cytotoxic CD4+ T cells in the lungs in severe COVID-19. Clinical Immunology (Orlando, Fla.), 237, 108991.
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2

Multiparametric Immunohistochemical Analysis

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Tissue samples were fixed in formalin, embedded in paraffin, and sectioned. These specimens were incubated with antibodies: CD3 (clone: A045229–2; DAKO), CD4 (clone: CM153A; Biocare), CD19 (clone: CM310 A, B; Biocare), CD8 (clone: ab85792; Abcam), CD31 (clone: 3528; Cell Signaling Technology), CD68 (clone: ab955; Abcam), Cleaved caspase-3 (clone: 9664; Cell Signaling Technology), GZMA (clone: LS-C312742; LSBio), CD28 (clone: ab243228; Abcam), CD45RA (clone: 158–4D3; Novus Biologicals), vimentin (clone: 5741; Cell Signaling Technology), α-SMA (clone: 19245; Cell Signaling Technology), pan-CK (clone: ab27988; Abcam), AQP5(clone: ab92320; Abcam), HLA-DR (clone: ab20181; Abcam) and SLAMF7 (clone: HPA055945; Sigma Aldrich) followed by incubation with secondary antibody using an OpalTM Multiplex Kit (Perkin Elmer). The samples were mounted with ProLong™ Diamond Antifade mountant containing DAPI (Invitrogen).
Perugino C.A., Kaneko N., Maehara T., Mattoo H., Kers J., Allard-Chamard H., Mahajan V.S., Liu H., Della-Torre E., Murphy S.J., Ghebremichael M., Wallace Z.S., Bolster M.B., Harvey L.M., Mylvaganam G., Tuncay Y., Liang L., Montesi S.B., Zhang X., Tinju A., Mochizuki K., Munemura R., Sakamoto M., Moriyama M., Nakamura S., Yosef N., Stone J.H, & Pillai S. (2020). CD4+ and CD8+ cytotoxic T lymphocytes may induce mesenchymal cell apoptosis in IgG4-related disease. The Journal of allergy and clinical immunology, 147(1), 368-382.
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3

Multiparametric Immunohistochemistry Profiling

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Tissue samples were fixed in formalin, embedded in paraffin, and sectioned. These specimens were incubated with antibodies: CD4 (clone: CM153A; Biocare), GATA3 (clone: CM405A; Biocare) and GZMA (clone: LS-C312742; LSBio) followed by incubation with secondary antibody using an OpalTM Multiplex Kit (Perkin Elmer). The samples were mounted with ProLong™ Diamond Antifade mountant containing DAPI (Invitrogen).
Allard-Chamard H., Alsufyani F., Kaneko N., Xing K., Perugino C., Mahajan V.S., Wheat J.L., Deepe GS J.r., Loyd J, & Pillai S. (2020). Cytotoxic CD4+ T cells in Fibrosing Mediastinitis linked to Histoplasma capsulatum. Journal of immunology (Baltimore, Md. : 1950), 206(3), 524-530.
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4

Multiplex Immunohistochemistry for Immune Cell Profiling

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Tissue samples were fixed in formalin, embedded in paraffin, and sectioned. These specimens were incubated with the following antibodies: anti-CD3 (clone: A045229-2; DAKO), anti-CD4 (clone: EPR6855;Abcam), anti-CD19 (clone: SKU310; Biocare Medical), anti-Bcl6 (clone: LN22; Biocare Medical), anti-AID (clone: ZA001; Invitrogen), anti-T-bet (clone: ab150440; Abcam), GATA3 (clone: CM405A; Biocare), ICOS (clone: 89601; Cell Signaling Technology), Rorc (clone: ab212496; Abcam), CXCR5 (clone: MAB190; R&D Systems), Foxp3 (clone: 98377; Cell Signaling Technology), anti-CD8 (clone: ab85792; Abcam), anti-IgD (clone: AA093; DAKO), anti-CD27 (clone: ab131254; Abcam), anti-IgG (clone: ab109489; Abcam), anti-TNF-α (clone: ab6671; Abcam), and anti-CD35 (clone: ab25; Abcam) followed by incubation with a secondary antibody using an Opal Multiplex Kit (Perkin Elmer). The samples were mounted with ProLong Diamond Antifade mountant containing DAPI (Invitrogen).
Kaneko N., Kuo H.H., Boucau J., Farmer J.R., Allard-Chamard H., Mahajan V.S., Piechocka-Trocha A., Lefteri K., Osborn M., Bals J., Bartsch Y.C., Bonheur N., Caradonna T.M., Chevalier J., Chowdhury F., Diefenbach T.J., Einkauf K., Fallon J., Feldman J., Finn K.K., Garcia-Broncano P., Hartana C.A., Hauser B.M., Jiang C., Kaplonek P., Karpell M., Koscher E.C., Lian X., Liu H., Liu J., Ly N.L., Michell A.R., Rassadkina Y., Seiger K., Sessa L., Shin S., Singh N., Sun W., Sun X., Ticheli H.J., Waring M.T., Zhu A.L., Alter G., Li J.Z., Lingwood D., Schmidt A.G., Lichterfeld M., Walker B.D., Yu X.G., Padera RF J.r, & Pillai S. (2020). Loss of Bcl-6-Expressing T Follicular Helper Cells and Germinal Centers in COVID-19. Cell, 183(1), 143-157.e13.
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5

Multiplex Immunohistochemistry of Immune Markers

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Tissue samples were fixed in formalin, embedded in paraffin, and sectioned. These specimens were incubated with the following antibodies: anti-CD4 (clone: CM153A; Biocare Medical), anti-CD19 (clone: SKU310; Biocare Medical), anti-CD20 (clone: LN26; Novus Biological), anti-Bcl6 (clone: LN22; Biocare Medical), anti-IgD (clone: AA093; DAKO), anti-CD27 (clone: ab131254; Abcam), anti-IgG (clone: ab109489; Abcam), anti-SLAMF7 (clone: HPA055945; Sigma-Aldrich) and anti-CD11c (clone: ab52632; Abcam) followed by incubation with a secondary antibody using an Opal™ Multiplex Kit (Perkin Elmer). The samples were mounted with ProLong™ Diamond Antifade mountant containing DAPI (Invitrogen).
Allard-Chamard H., Kaneko N., Bertocchi A., Sun N., Boucau J., Kuo H.H., Farmer J.R., Perugino C., Mahajan V.S., Murphy S.J., Premo K., Diefenbach T., Ghebremichael M., Yuen G., Kotta A., Akman Z., Lichterfeld M., Walker B.D., Yu X.G., Moriyama M., Maehara T., Nakamura S., Stone J.H., Padera R.F, & Pillai S. (2023). Extrafollicular IgD−CD27−CXCR5−CD11c− DN3 B cells infiltrate inflamed tissues in autoimmune fibrosis and in severe COVID-19. Cell Reports, 42(6), 112630.
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6

Multiplex IF Profiling of Prostate Tissues

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5 μm-thick formalin-fixed paraffin-embedded sections from tissue microarray (PR484a: 24 cases/48 cores from US BIOMAX) were stained using Opal multiplex kits, according to the manufacturer’s protocol (PerkinElmer, Waltham, MA, USA), for a panel of DAPI, and GNRH2. Multiplex immunofluorescence scans were captured by a PerkinElmer Vectra Polaris. Mean fluorescence intensity was measured in prostate epithelium (benign or malignant) using open source QuPath v 0.2.0 (stable version).
SISSUNG T.M., LOCHRIN S., LIU T., SCHMIDT K., STROPE J., RISDON E., CHOO-WOSOBA H., VENZON D.J., LASSOUED W., SATER H.A., WALTER-RODRIGUEZ B., PRICE D.K, & FIGG W.D. (2023). GNRH2 Polymorphism in Men With Prostate Cancer Treated With Androgen Deprivation Therapy. Anticancer research, 43(9), 4023-4030.
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7

Multiplex Imaging Analysis of Immune Cells in HPV-Negative HNSCC Tumors

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Tumor biopsies from advanced stage (III/IV) patients with human papillomavirus (HPV)-negative HNSCC were obtained at the time of standard-of-care surgical resection under institutional review board-approved protocol NCT03429036 with informed consent. Sections were stained using Opal multiplex kits, according to the manufacturer’s protocol (PerkinElmer), for a panel of 4′,6-diamidino-2-phenylindole (DAPI), pan-cytokeratin (clone AE1/AE3), HLA class I (EMR8-5) and CD3 (SP7). Positive and negative controls included normal human tonsil and unstained sections (no primary antibody), respectively. After optimization, staining of all analyzed sections was done in a single automated assay. Images were captured by a PerkinElmer Vectra Polaris. Multispectral fluorescent images were analyzed using QuPath software.20 (link) Following annotation of entire tumor sections into tumor and stromal tissue based on cytokeratin staining, HLA positivity and CD3+ cells were quantified using automated detection based on universally applied fluorescence detection thresholds.
Lee M.Y., Robbins Y., Sievers C., Friedman J., Abdul Sater H., Clavijo P.E., Judd N., Tsong E., Silvin C., Soon-Shiong P., Padget M.R., Schlom J., Hodge J., Hinrichs C, & Allen C. (2021). Chimeric antigen receptor engineered NK cellular immunotherapy overcomes the selection of T-cell escape variant cancer cells. Journal for Immunotherapy of Cancer, 9(3), e002128.
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8

Multiplex Immunohistochemistry for Immune Cell Profiling

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Tumor specimens were stained using Opal Multiplex Kits, according to the manufacturer's protocol (Perkin-Elmer), for CD8 (clone 4B11; Leica, PA0183), CD68 (clone KP1; BioGenex, Am416-5M), HLA-DR (EP96; BioSB, BSB6797), CD163 (clone OTI2G1; Abcam, ab156769), CD206 (clone 15-2; Abcam, ab64693), and CSF1R (clone EPR20754; Abcam, ab229188). Specimens underwent serial staining with primary and secondary antibodies followed by the use of a covalently bound tyramide signal amplification process (13, 14 (link)).
Manji G.A., Van Tine B.A., Lee S.M., Raufi A.G., Pellicciotta I., Hirbe A.C., Pradhan J., Chen A., Rabadan R, & Schwartz G.K. (2021). A Phase I Study of the Combination of Pexidartinib and Sirolimus to Target Tumor-Associated Macrophages in Unresectable Sarcoma and Malignant Peripheral Nerve Sheath Tumors. Clinical Cancer Research, 27(20), 5519-5527.
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