Envision detection kit

Manufactured by Agilent Technologies

Sourced in Denmark, United States, Germany, China

The EnVision detection kit is a laboratory instrument designed to detect and quantify a variety of analytes in samples. It provides a sensitive and reliable platform for various assays, including fluorescence, luminescence, and absorbance measurements.

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Lab products found in correlation

Transforming growth factor 1, by Thermo Fisher Scientific (4 mentions) Rabbit antihuman collagen type 2, by Abcam (4 mentions) Sodium pyruvate, by Thermo Fisher Scientific (4 mentions) Insulin transferrin selenium, by Thermo Fisher Scientific (4 mentions) Dexamethasone (dex), by Merck Group (4 mentions) Ascorbic acid, by Merck Group (4 mentions) Photomicroscope, by Olympus (3 mentions) Image pro plus 6, by Media Cybernetics (3 mentions) Envision flex target retrieval solution, by Agilent Technologies (2 mentions) Target retrieval solution, by Agilent Technologies (2 mentions) Diaminobenzidine substrate kit, by Agilent Technologies (2 mentions) Scanscope scanner, by Leica (2 mentions) Fgfr2 antibody, by Abcam (1 mentions) P erk, by Cell Signaling Technology (1 mentions) P akt, by Cell Signaling Technology (1 mentions) Clone ep51, by Agilent Technologies (1 mentions) Cd147 antibody, by Santa Cruz Biotechnology (1 mentions) Light microscopy, by Olympus (1 mentions) Normal goat serum (ngs), by Abcam (1 mentions) Ab16667, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Envision kit (439 citations) ChemMate DAKO EnVision Detection Kit (27 citations) EnVision Detection System kit (23 citations) EnVision FLEX Detection Kit (21 citations) Dako REAL EnVision Detection System kit (7 citations) Envision Plus Detection Kit (5 citations) ChemMate TM EnVision System/DAB Detection Kits (3 citations) Envision DAB detection kit (2 citations) Dako EnVision+System HRP labelled polymer detection kit (2 citations) EnVision + System-HRP (DAB) detection kit (2 citations)

91 protocols using envision detection kit

Immunohistochemical Analysis of FGFR2 and MAPK Signaling

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Representative 4-μm serial sections of the tumor were prepared from 10% FFPE tissue blocks for immunohistochemistry. Briefly, all slides were exposed to 3% hydrogen peroxide for 10 min to block endogenous peroxidase activity. FGFR2 antibody (#AB58201, 1:300, anti-mouse, Abcam, USA), ERK (#4695, 1:250, anti-rabbit, Cell signaling technology, USA), P-ERK (#4370, 1:250, anti-rabbit, Cell signaling technology, USA), AKT (#4685, 1:200, anti-rabbit, Cell signaling technology, USA), P-AKT (#4060, 1:100, anti-rabbit, Cell signaling technology, USA) incubated with tumor sections in a humidified chamber at 4 °C overnight, followed by the secondary anti-mouse peroxidase-conjugated secondary antibody (EnVision^TM Detection Kit, DAKO, Denmark) or anti-rabbit peroxidase-conjugated secondary antibody (EnVision^TM Detection Kit, DAKO, Denmark) at 37 °C for 30 min.
The IHC score was calculated by multiplying the staining intensity (0 = no staining, 1 = mild staining, 2 = moderate staining, and 3 = strong staining) by the percentage of immunoreactive tumor cells (0–100). The immunostaining result was considered to be 0 or negative when the score was <25; 1+ or weak when the score was 26–100; 2+ or moderate when the score was 101–200; or 3+ or strong when the score was 201–300.

Pu X., Ye Q., Cai J., Yang X., Fu Y., Fan X., Wu H., Chen J., Qiu Y, & Yue S. (2021). Typing FGFR2 translocation determines the response to targeted therapy of intrahepatic cholangiocarcinomas. Cell Death & Disease, 12(3), 256.

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Immunohistochemical Evaluation of Mismatch Repair Proteins

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Representative tumor areas were selected by experienced pathologists, evaluated in hematoxylin-and-eosin (H&E) sections, and employed for TMA inclusion selection, as previously described [46 (link)]. MMR status was determined using IHC with specific primary antibodies for MLH1 (clone ES05, #IR079), PMS2 (clone EP51, #IR087), MSH2 (clone FE11, #IR085), and MSH6 (clone EP49, #IR086), and with the Envision detection kit (all from Agilent Technologies, Santa Clara, CA, USA). Entire sections from the original tissue block were examined when the case was not assessable in the TMA.
The presence of dMMR was considered when the complete loss of nuclear expression of one or more markers in tumor cells was observed. Normal stromal tissue cells demonstrating nuclear staining served as an internal positive control [47 (link)]. Additionally, MSH3 expression was analyzed with a primary specific antibody (clone RM405, #275928, Abcam, Cambridge, UK).

Mendiola M., Heredia-Soto V., Ruz-Caracuel I., Baillo A., Ramon-Patino J.L., Escudero F.J., Miguel M., Pelaez-Garcia A., Hernandez A., Feliu J., Hardisson D, & Redondo A. (2023). Comparison of Methods for Testing Mismatch Repair Status in Endometrial Cancer. International Journal of Molecular Sciences, 24(19), 14468.

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Immunohistochemical Analysis of Sirt7 Expression

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The archived paraffin- embedded tissues were cut to a thickness of 4 µm. Immunostaining was performed using the two-step Elivision Plus kit system (Fuzhou Maixin Biotech Co., Ltd., Fuzhou, China). The sections were dewaxed in xylene, rehydrated with a series of ethanol solutions (100, 95, 80 and 70%), and then boiled in citrate buffer (pH 6.0) for 2 min in an autoclave. Next, 0.3% H₂O₂ was used to block endogenous peroxidase activity at room temperature for 15 min, and the sections were incubated with normal goat serum (dilution ratio 1:20) for 20 min at room temperature to reduce non-specific binding. Tissue sections were incubated with the Sirt7 antibody (cat. no. 5360s; 1:150 dilution; Santa Cruz Biotechnology, Inc.) for 2 h at room temperature. The secondary antibody was applied using the Envision Detection kit (Dako; Agilent Technologies, Inc., Santa Clara, CA, USA). Slides were stained for 2 min with diaminobenzidine tetrahydrochloride (DAB) and then counterstained 2 min with hematoxylin at room temperature. The stained tumor cells were assessed with a Nikon microscope in 10 independent fields at magnification, ×400.

Mu P., Liu K., Lin Q., Yang W., Liu D., Lin Z., Shao W, & Ji T. (2018). Sirtuin 7 promotes glioma proliferation and invasion through activation of the ERK/STAT3 signaling pathway. Oncology Letters, 17(2), 1445-1452.

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Immunohistochemical Analysis of BI-ALCL

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Immunohistochemistry for GATA3 (clone L50-823, Cell Marque, Rocklin, CA) and FoxP3 (clone 236A/E7, Abcam, UK) was performed on 2 µm thick formalin-fixed paraffin-embedded (FFPE) tissue sections of a BI-ALCL seroma cell block and of a BI-ALCL xenograft using an automated immunostainer (Omnis, Agilent Technologies, USA). Tissue sections were pretreated using EnVisionTM FLEX Target Retrieval Solution (Agilent) and incubated with an optimal dilution of the primary antibody. The reaction was visualized with the EnVision Detection Kit (Agilent) using 3–3′-diaminobenzidine chromogenic substrate. Sections were counterstained with EnVision FLEX Hematoxylin (Link) (Agilent).

Di Napoli A., Greco D., Scafetta G., Ascenzi F., Gulino A., Aurisicchio L., Santanelli Di Pompeo F., Bonifacino A., Giarnieri E., Morgan J., Mancini R, & Kadin M.E. (2020). IL-10, IL-13, Eotaxin and IL-10/IL-6 ratio distinguish breast implant-associated anaplastic large-cell lymphoma from all types of benign late seromas. Cancer Immunology, Immunotherapy, 70(5), 1379-1392.

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Immunohistochemical Analysis of Inflammatory Markers

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Sections were treated with 3% (v/v) H₂O₂ in methanol to block endogenous peroxidase activity. Immunohistochemistry was performed using the Envision Detection™ kit (DAKO Agilent Technologies Inc., Santa Clara, CA, USA). Tissue sections were incubated with primary antibodies against IL-6, TNF-α, IL-1β, IL-17, phosphorylated (p) STAT3 (Tyr705), and TRAP for 2 h at room temperature. Sections were then incubated with a biotinylated secondary Ab and streptavidin–peroxidase complex for 30 min. The final colored products were developed using chromogen diaminobenzidine, and the sections were examined under a photomicroscope (Olympus, Tokyo, Japan). Two independent, blinded observers assessed the number of positive cells per section at high-power field (magnifications ×400). To analyze osteoclast parameters, osteoclast surface normalized to the bone surface were assessed by bone histomorphometric analyses (ImageJ software).

Park J.S., Yang S.C., Jeong H.Y., Lee S.Y., Ryu J.G., Choi J.W., Kang H.Y., Kim S.M., Hwang S.H., Cho M.L, & Park S.H. (2022). EC-18 prevents autoimmune arthritis by suppressing inflammatory cytokines and osteoclastogenesis. Arthritis Research & Therapy, 24, 254.

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Immunohistochemical Analysis of Cyclophilin A and CD147

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Immunohistochemistry was performed in 4 μm paraffin sections. The tissue sections were dewaxed in xylene, rehydrated in graded ethanol, and rinsed in distilled water. Citrate buffer (pH = 6.0) was used to retrive the antigen, and 0.3% H₂O₂ was used to block endogenous peroxidase activity. After the blockade of nonspecific binding, tissue sections were incubated with the Cyclophilin A antibody (cat. no. 5360s; 1 : 150 dilution; Santa Cruz Biotechnology, Inc.) and CD147 antibody (cat. no. 13287; 1 : 400 dilution; Cell Signaling Technology, Inc.) for overnight at 4 centigrade. The secondary antibody was applied using the Envision Detection kit (Dako; Agilent Technologies, Inc., Santa Clara, CA, USA). The staining was performed by the application of diaminobenzidine tetrahydrochloride (DAB) for 2 min and hematoxylin for 1 min at room temperature. The stained sections were evaluated with a Nikon microscope in 10 independent fields at magnification, ×400. We followed the methods of Mu et al. [20 (link)].

Xu S., Hu C., Xiao Z., Luo C, & Liu Z. (2020). Downregulation of CyclophilinA/CD147 Axis Induces Cell Apoptosis and Inhibits Glioma Aggressiveness. BioMed Research International, 2020, 7035847.

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Immunohistochemical Analysis of Inflammatory Markers

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Sections were treated with 3% (v/v) H₂O₂ in methanol to block endogenous peroxidase activity. Immunohistochemistry was performed using the Envision Detection™ kit (DAKO Agilent Technologies Inc., Santa Clara, CA, USA). Tissue sections were incubated with primary antibodies against IL-1β, TNF-α, and VEGF for 2 h at room temperature. Sections were then incubated with a biotinylated secondary antibody and streptavidin–peroxidase complex for 30 min. The final colored products were developed using chromogen diaminobenzidine. The sections were examined by light microscopy (Olympus, Tokyo, Japan). The number of positive cells was counted at high-power field (magnifications × 400) with the aid of Adobe Photoshop software by four individuals and averaged three randomly selected fields per tissue section.

Park J.S., Lee D., Yang S., Jeong H.Y., Na H.S., Cho K.H., Choi J., Koo H., Cho M.L, & Park S.H. (2022). Methotrexate-loaded nanoparticles ameliorate experimental model of autoimmune arthritis by regulating the balance of interleukin-17-producing T cells and regulatory T cells. Journal of Translational Medicine, 20, 85.

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Immunohistochemistry Protocol for Ki-67 and γ-H2AX

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Tissues were fixed with 10% buffered formalin at 4°C for overnight and embedded in paraffin (FFPE). FFPE were sectioned at 4-μm thickness. For IHC staining, tissue sections were deparaffinized and rehydrated, endogenous peroxidase activity was quenched by incubating the sections in a 3% hydrogen peroxide (H2O2)-methanol solution for 30 min at room temperature. Antigen retrieval was then subjected to 0.01 M citrate buffer (pH 6.0) for 13 min at high temperature (95-100°C). Tissues were blocked with 10% normal goat serum (Abcam, Cambridge, UK) in phosphate-buffered saline (PBS) for 20 min at room temperature. Primary antibodies against Ki-67 (Abcam, ab16667) and γ-H2AX (Cell Signaling, #9718) were used for staining following dilution in antibody diluent. HRP-conjugated secondary antibodies were used, and followed by DAB Chromogen staining using rabbit/mouse peroxidase/3, 3’-diaminobenzidine (DAB) EnVision™ Detection kit (Agilent-Dako, Califorlia, UAS). Tissue sections were counterstained with hematoxylin to show cell nuclei. Five fields per tumor section were quantified using Image J software for Ki-67 or γ-H2AX-positive cell staining with a minimum sample size of 5 animals per cohort.

Xu C., Gao Q., Wu Z., Lou W., Li X., Wang M., Wang N, & Li Q. (2022). Combined HASPIN and mTOR inhibition is synergistic against KRAS-driven carcinomas. Translational Oncology, 26, 101540.

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Immunohistochemical Analysis of UBE2M

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Part of the collected CRC and normal tissues were fixed with formalin and embedded in paraffin. The embedded tissues were then cut into 5 µm-thick sections, mounted onto slides, and then subjected to immunohistochemical staining, as described previously (16 (link)). Briefly, the slides were dehydrated in graded alcohol and endogenous peroxidase was removed with 3% hydrogen peroxide for 10 min. The slides were then blocked using 5% goat serum for 30 min and incubated with rabbit polyclonal anti-UBE2M antibody at a 1:200 dilution for 60 min. Next, the slides were incubated with horseradish peroxidase-conjugated secondary antibody (Dako, USA) at a 1:100 dilution for 60 min. Finally, the slides were stained using the EnVisionTM Detection Kit (Dako, USA), followed by hematoxylin staining.

Xu J., Lv G., Xu B, & Jiang B. (2020). Overexpression of UBE2M through Wnt/β-Catenin signaling is associated with poor prognosis and chemotherapy resistance in colorectal cancer. Translational Cancer Research, 9(9), 5614-5625.

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Immunohistochemical Analysis of Lymphatic Markers

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Briefly, serial 4 μm sections were deparaffinized by incubation in xylene, at 60°C, and rehydrated in a series of graded alcohol solutions, followed by washing in tris-buffered saline (TBS) (pH 7.6). Endogenous peroxidase activity was blocked with 0.3% hydrogen peroxide in distilled water, followed by washing in TBS. The sections were treated in a microwave oven, in a citrate buffer (pH 6.0), for antigen retrieval. Non-specific binding was blocked by treating the slides for 12 minutes with 2% bovine serum albumin in TBS. The commercially available antibodies for podoplanin (D2-40), LYVE-1, and VEGF-C were used as described in Table 1. The bound primary antibody was detected with the Envision TM detection kit (DAKO, Hamburg, Germany) and visualized using diaminobenzidine (DAB) as the chromogen. Finally, the tissue sections were counterstained with hematoxylin and dehydrated through graded ethanols and xylene.

Kostis G., Ioannis L., Helen K, & Helen P. (2014). The expression of vascular endothelial growth factor-C correlates with lymphatic microvessel density and lymph node metastasis in prostate carcinoma: An immunohistochemical study. Urology Annals, 6(3), 224-230.

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