Ultravision detection system

Manufactured by Thermo Fisher Scientific

Sourced in United States

The UltraVision Detection System is a laboratory equipment product designed for the detection and visualization of target proteins in biological samples. It provides a reliable and sensitive method for immunohistochemistry (IHC) and in situ hybridization (ISH) applications. The system includes all the necessary components to perform these analyses, such as reagents, detection kits, and protocols.

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Lab products found in correlation

Histofine sab po kit, by Nichirei Biosciences (3 mentions) Ab92742, by Abcam (2 mentions) Hpa008572, by Merck Group (2 mentions) Ab185954, by Abcam (2 mentions) Ab9474, by Abcam (1 mentions) Desmin, by ABclonal (1 mentions) A19040, by ABclonal (1 mentions) Gadd45g, by ABclonal (1 mentions) Ba2879, by Boster Bio (1 mentions) Ab5694, by Abcam (1 mentions) Ab51608, by Abcam (1 mentions) Hpa021241, by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by LGC (1 mentions) Bs 0061r, by Bioss Antibodies (1 mentions) Fluoromount, by Diagnostic BioSystems (1 mentions) Ab150077, by Abcam (1 mentions) Gapdh, by Merck Group (1 mentions) Mab374, by Merck Group (1 mentions) Phenol red, by Merck Group (1 mentions) Fibronectin, by Merck Group (1 mentions)

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32 protocols using ultravision detection system

Immunohistochemistry of Lung KI67 Expression

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IHC was performed on mouse lung tissue to examine KI67 expression. Lung sections were deparaffinized and rehydrate to water. Antigens were retrieved by heating the sections in BorgDecloaketRTU (Biocare medical, Concord, CA) for 10 minutes at 95C. After antigen retrieval, non-specific binding was blocked using Ultra V Block (UltraVision Detection System, Thermoscientific, Fremont, CA) for 10 minutes, then slides were incubated in humidity chamber with primary antibody from ABCAM (Cambridge, MA) at 1:4000 dilution for 1 hour at room temperature. As negative controls, adjacent slides were incubated in normal rabbit serum. After two buffer washes, slides were incubated for 10 minutes with biotinylated Goat Anti-polyvalent (UltraVision Detection System, Thermo Scientific), followed by a buffer wash and incubated for 10 minute in Streptavidin Peroxidase (UltraVision Detection System from Thermo scientific, Fremont, CA). After a buffer wash, staining of interest was developed with DAB chromagen (Dako, Carpinteria, CA) and counterstained with Fast Green (Sigma St. Louis, MO). Slides were then dehydrated and mounted with Cytoseal (Thermo Scientific, Fremont, CA). Quantification of immunohistochemistry staining was done as previously described.[11 (link)]

McGrath-Morrow S.A., Hayashi M., Aherrera A., Lopez A., Malinina A., Collaco J.M., Neptune E., Klein J.D., Winickoff J.P., Breysse P., Lazarus P, & Chen G. (2015). The Effects of Electronic Cigarette Emissions on Systemic Cotinine Levels, Weight and Postnatal Lung Growth in Neonatal Mice. PLoS ONE, 10(2), e0118344.

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Immunostaining of Prostate Cancer Specimens

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A total of eight CRPC specimens were used (Supplementary Table 2). Tissue specimens were immunostained following the manufacturer's protocol with the Ultra-Vision Detection System (Thermo Scientific). Primary rabbit polyclonal antibodies against AR (1 : 50, ab9474; Abcam), antibodies against PSA (1 : 500, HPA000764; Sigma-Aldrich), and antibodies against Ecm29 (1 : 500, PA5-29467; Pierce Antibodies, Thermo Scientific) were used for immunochemistry. The slides were treated with biotinylated goat antibodies (Histofine SAB-PO kit; Nichirei, Tokyo, Japan).

Goto Y., Kojima S., Nishikawa R., Kurozumi A., Kato M., Enokida H., Matsushita R., Yamazaki K., Ishida Y., Nakagawa M., Naya Y., Ichikawa T, & Seki N. (2015). MicroRNA expression signature of castration-resistant prostate cancer: the microRNA-221/222 cluster functions as a tumour suppressor and disease progression marker. British Journal of Cancer, 113(7), 1055-1065.

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Comprehensive Histological Analysis of Pancreatic Tissue

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Pancreatic tissues were fixed in 10% formalin solution, embedded in paraffin, and sectioned in 3-μm slices. Sections were stained with H&E and Masson’s trichrome to detect the interstitial volume expansion. Antigen retrieval was performed in sodium citrate buffer at 96°C. Sections were washed with PBS three times, and endogenous peroxidase activity was blocked by incubating sections in a solution of 3% H₂O₂ for 5 min. Sections were then blocked with 10% normal goat serum. The primary antibodies used were α-SMA (ab5694, Abcam), Desmin (A3736, Abclonal), GFAP (A19058, Abclonal), KRT19 (A19040, Abclonal), Gadd45g (A10286, Abclonal), and Tsc1 (BA2879, BOSTER). Incubation with the primary antibody was performed at 4°C overnight. Tissue sections were developed using the Ultra Vision Detection System (Thermo Fisher Scientific).

Li F., Wang M., Li X., Long Y., Chen K., Wang X., Zhong M., Cheng W., Tian X., Wang P., Ji M, & Ma X. (2022). Inflammatory-miR-301a circuitry drives mTOR and Stat3-dependent PSC activation in chronic pancreatitis and PanIN. Molecular Therapy. Nucleic Acids, 27, 970-982.

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Immunoblotting and Immunohistochemistry Protocols

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Immunoblotting was carried out as previously described (15 (link)) with diluted (1:1000) anti-PHGDH antibodies (HPA021241; Sigma), anti-HIF2α antibodies (ab51608; Abcam, Cambridge, MA, USA), anti-glyceraldehyde 3-phosphate dehydrogenase antibodies (GAPDH; MAB374, EMD Millipore, Billerica, MA), and anti-β-actin antibodies (bs-0061R; Bioss, Woburn, MA, USA). Immunohistochemistry were performed using an UltraVision Detection System (Thermo Scientific, Fremont, CA, USA) according to the manufacturer’s instructions. The primary rabbit monoclonal antibodies against Ki67 (ab92742; Abcam) were diluted 1:100. For immunofluorescence analyses, nuclei were stained with DAPI (1 μg/mL; Kirkegaard & Perry Laboratories, Gaithersburg, MD, USA), and slides were mounted in Fluoromount (Diagnostic Biosystems, Pleasanton, CA, USA). Anti-PHGDH antibodies (HPA021241; Sigma) were used as the primary antibody at a dilution of 1:100, and binding was visualised using secondary antibodies conjugated to Alexa Fluor 488 (ab150077; Abcam).

Yoshino H., Nohata N., Miyamoto K., Yonemori M., Sakaguchi T., Sugita S., Itesako T., Kofuji S., Nakagawa M., Dahiya R, & Enokida H. (2017). PHGDH as a key enzyme for serine biosynthesis in HIF2α-targeting therapy for renal cell carcinoma. Cancer research, 77(22), 6321-6329.

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Immunohistochemical Profiling of Aortic ECM

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Native and decellularised zinc fixed and paraffin embedded tissue sections (from n = 6 human aortas) were stained with monoclonal antibodies against collagen I (Millipore; IgG isotype, polyclonal, 1:500 dilution), collagen III (Millipore; IgG1 isotype, clone IE7-D7, 1:100 dilution), collagen IV (Dako; IgG1 isotype, clone PHM-12, 1:200 dilution), von Willebrand factor (Dako; IgG isotype, polyclonal, 1:400 dilution), fibronectin (Vector; IgG1 isotype, clone 568, 1:150 dilution) and laminin (Sigma; IgG1 isotype, clone LAM-89, 1:1000 dilution). Immunolabeling was carried out using an Ultravision detection system (Thermo Scientific). Antigen retrieval was employed using proteinase K (Dako; room temperature for 20 min—collagen III, von Willebrand factor, fibronectin and laminin), proteinase K followed by trypsin digestion (0.05% w/v trypsin 0.52 mM EDTA.4Na in HBSS with phenol red (Sigma) for 30 s) (collagen IV) and Vector antigen unmasking solution citric based (collagen I). Hydrogen peroxide (Sigma) was used to block endogenous peroxidase activity. Isotype control antibodies (IgG and IgG1; Dako; used at the same concentrations as the test antibody) and omission of the primary antibody served as negative controls. Sections were viewed using Kohler and all images were captured digitally using Zen Blue Pro (Zeiss).

Aldridge A., Desai A., Owston H., Jennings L.M., Fisher J., Rooney P., Kearney J.N., Ingham E, & Wilshaw S.P. (2017). Development and characterisation of a large diameter decellularised vascular allograft. Cell and Tissue Banking, 19(3), 287-300.

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Immunohistochemical Analysis of ITGA3 in Breast Cancer

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A tissue microarray of BC samples was obtained from US Biomax Inc. (product ID: BL 1002; Rockville, MD, USA). Detailed information on all tumour specimens can be found at http://www.biomax.us/index.php. The tissue microarray was immunostained following the manufacturer's protocol with an Ultra Vision Detection System (Thermo Scientific). Primary rabbit polyclonal antibodies against ITGA3 (HPA008572; Sigma-Aldrich) were diluted 1 : 20. Immunostaining was evaluated according to the scoring method described previously (Yoshino et al, 2011 (link)). Each case was scored on the basis of the intensity and area of staining. The intensity of staining was graded on the following scale: 0, no staining; 1+, mild staining; 2+, moderate staining; and 3+, intense staining. The area of staining was evaluated as follows: 0, no staining of cells in any microscopic field; 1+,<30% of cells stained positive; 2+, 30–60% stained positive; 3+,>60% stained positive. The immunostaining scores (intensity+extent) were combined and analysed. All samples were independently scored by two of the authors (T Sakaguchi and K Miyamoto), who were blinded to the patient status.

Sakaguchi T., Yoshino H., Yonemori M., Miyamoto K., Sugita S., Matsushita R., Itesako T., Tatarano S., Nakagawa M, & Enokida H. (2017). Regulation of ITGA3 by the dual-stranded microRNA-199 family as a potential prognostic marker in bladder cancer. British Journal of Cancer, 116(8), 1077-1087.

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Immunohistochemistry for Ki67 Expression

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Immunohistochemistry was performed using an UltraVision Detection System (Thermo Scientific, Fremont, CA, USA) according to the manufacturer's instructions. The primary rabbit monoclonal antibodies against Ki67 (ab92742; Abcam, Cambridge, UK) were diluted 1 : 500 and incubated at 4 °C overnight. The secondary antibody was Goat Anti‐Rabbit IgG Antibody (H + L), Biotinylated (BA‐1000; Vector Laboratories, San Francisco, CA, USA) diluted to 5 µg·mL⁻¹ and incubated for 30 min. Positive cells were quantitated by counting six random microscopic fields. These experimental procedures were described in a previous report [27 (link)].

Tamai M., Tatarano S., Okamura S., Fukumoto W., Kawakami I., Osako Y., Sakaguchi T., Sugita S., Yonemori M., Yamada Y., Nakagawa M., Enokida H, & Yoshino H. (2022). microRNA‐99a‐5p induces cellular senescence in gemcitabine‐resistant bladder cancer by targeting SMARCD1. Molecular Oncology, 16(6), 1329-1346.

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Immune Profiling of Prostate Tumors

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Pre- and post-chemotherapy plasma vascular endothelial growth factor (VEGF) were measured. Tumor expression of tissue VEGF was characterized by tissue microarray. Methods for these assays have been described [20 (link)]. Also, serum total testosterone levels were obtained prior to and after neoadjuvant chemotherapy.
Immune infiltration was examined via immunohistochemistry with a panel of 8 markers in primary prostatic tumors and adjacent normal tissue using a tissue microarray (TMA) of 50 patients treated neoadjuvantly with docetaxel and mitoxantrone. Heat mediated antigen retrieval was performed with citrate buffer (BioGenex, Fremont, CA), with the exception of retrieval prior to CD4 staining, which was achieved using 10 mM Tris/1 mM EDTA. Staining was performed with the Ultravision Detection System (Thermo Scientific, Waltham, MA), followed by counterstaining with methyl green. Antibodies CD3, CD4, CD8, CD15, and CD163 were from Thermo Scientific, CD68 (clone PG-M1) and CD20 were from AbCAM (Cambridge, UK), and FoxP3 was from eBioscience (San Diego, CA). Slides were digitally scanned with an Aperio ScanScope CT Slide Scanner at 40x objective and individual TMA cores were analyzed separately using TMALab (Aperio, Buffalo Grove, IL) and the nuclear stain detection algorithm to quantify the number of positive cells within each core. Data is displayed as cell density (cells/mm²).

Bergstrom C.P., Ruffell B., Ho C.M., Higano C.S., Ellis W.J., Garzotto M., Beer T.M, & Graff J.N. (2017). Docetaxel and Mitoxantrone Prior to Radical Prostatectomy in Men with High Risk Prostate Cancer: Ten-Year Follow Up and Immune Correlates. Anti-cancer drugs, 28(1), 120-126.

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Quantifying Apoptosis in Testis

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Sodium citrate buffer (0.01 M and pH 6) was used on testis sections with an antigen retrieval step. UltraVision detection system (Thermo-Scientific, USA) was applied in immunohistochemistry assays, as previously described by Urriola-Muñoz, P. et al.^{22 (link)}. Slides were incubated overnight at 4 °C with an anti-caspase 3 antibody (Cell Signaling, USA) at 1 mg/ml and the sections counter-stained with hematoxylin and subsequently evaluated under a microscope. Active caspase-3 cells were quantified in a minimum of 100 seminiferous tubules per each replicate, per (n).

Buñay J., Larriba E., Moreno R.D, & del Mazo J. (2017). Chronic low-dose exposure to a mixture of environmental endocrine disruptors induces microRNAs/isomiRs deregulation in mouse concomitant with intratesticular estradiol reduction. Scientific Reports, 7, 3373.

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Histological Analysis of Lung Tissues

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The lung tissues were fixed in 10% formalin solution. Paraffin was used to encapsulate the chips, and then 3-μm-thick slices were prepared. The sections were stained with H&E and Masson’s trichrome to detect interstitial volume expansion. The antigen was repaired with sodium citrate buffer in a water bath at 96°C. After washing with phosphate-buffered saline three times and 3% H₂O₂ for 5 min, the sections were blocked with 10% normal goat serum. For immunohistochemical staining to measure the levels of α-SMA, Ki67, Tsc1, and p-mTOR, the lung tissue sections were incubated with these primary antibodies at 4°C overnight. The tissue sections were developed using an Ultra Vision Detection System (Thermo Fisher Scientific, MA, USA).

Wang J., Li X., Zhong M., Wang Y., Zou L., Wang M., Gong X., Wang X., Zhou C., Ma X, & Liu M. (2020). miR-301a Suppression within Fibroblasts Limits the Progression of Fibrosis through the TSC1/mTOR Pathway. Molecular Therapy. Nucleic Acids, 21, 217-228.

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