Envision system hrp labeled polymer anti mouse

Manufactured by Agilent Technologies

Sourced in United States, Denmark

The Envision System-HRP Labeled Polymer Anti-Mouse is a laboratory equipment product designed for use in immunohistochemistry (IHC) procedures. It is a horseradish peroxidase (HRP)-labeled polymer that can be used to detect mouse primary antibodies. The product facilitates the visualization of target antigens in tissue sections or cell samples.

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Lab products found in correlation

Mayer s hematoxylin, by Merck Group (2 mentions) Lfmb 21, by Santa Cruz Biotechnology (2 mentions) 3 3 diaminobenzidine (dab), by Agilent Technologies (2 mentions) Osteosoft, by Merck Group (2 mentions) K3465, by Agilent Technologies (1 mentions) Bz h3c, by Keyence (1 mentions) Novolink polymer, by Leica (1 mentions) Dab substrate chromogen system, by Agilent Technologies (1 mentions) Mib 5, by Agilent Technologies (1 mentions) Target retrieval solution, by Agilent Technologies (1 mentions) Endogenous peroxidase block, by Agilent Technologies (1 mentions) Autostainer xl, by Leica (1 mentions) Herceptest, by Agilent Technologies (1 mentions) 3 3 diaminobenzidine tetrahydrochloride, by Merck Group (1 mentions)

Close spelling variants of this product

Dako Envision + System-HRP Labeled Polymer Anti-mouse Envision System HRP-labeled polymer anti-rabbit or anti-mouse secondary antibodies EnVision™ system-horseradish peroxidase (HRP)-labeled polymer anti-mouse EnVision + System-HRP Labeled Polymer anti-rabbit or anti-mouse EnVision+ System-HRP Labeled Polymer Anti-mouse Envision polymer Anti-mouse EnVision-HRP Labeled Polymer-HRP Envision®+system (anti-mouse) EnVision+ System-HRP

9 protocols using envision system hrp labeled polymer anti mouse

Ovarian CYP27B1 and Ki67 Expression

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The expression of CYP27B1 in ovarian tissues was detected using immunohistochemistry, as previously described (48 (link)). Briefly, formalin-fixed paraffin-embedded 4- to 5-μm sections were labeled overnight at 4°C with rabbit anti-CYP27B1 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at a dilution of 1:75. The antigen-antibody binding was visualized with HRP-labeled anti-rabbit antibody and 3,3′-diaminobenzidine (DAB) (Envision System-HRP Labeled Polymer Anti-Mouse; Dako, Glostrup, Denmark), followed by hematoxylin counterstaining. The kidney sections served as positive control.
Furthermore, Ki67 immunostaining was performed as described previously (21 (link),49 (link),50 (link)).

BROŻYNA A.A., JÓŹWICKI W., JOCHYMSKI C, & SLOMINSKI A.T. (2014). Decreased expression of CYP27B1 correlates with the increased aggressiveness of ovarian carcinomas. Oncology Reports, 33(2), 599-606.

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Immunohistochemical Analysis of HER3 Expression

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Xenograft tumors of HER3/RH7777, RH7777, and CTOS C45 were excised and fixed in neutralized 10% formalin, embedded in paraffin, and cut into 4-μm-thick sections. The sections were incubated with diluted mouse anti-HER3 monoclonal antibody (2.5 μg/mL; erbB3/Her3, nanoTools, Teningen, Germany) for 1 h at room temperature, washed, and then incubated with peroxidase-conjugated secondary antibodies (EnVision+ System-HRP Labeled Polymer Anti-mouse; DAKO, Glostrup, Denmark). The binding of the secondary antibodies was visualized with diaminobenzidine staining (K3465; DAKO) and counterstaining was carried out with hematoxylin.

Yuan Q., Furukawa T., Tashiro T., Okita K., Jin Z.H., Aung W., Sugyo A., Nagatsu K., Endo H., Tsuji A.B., Zhang M.R., Masuko T., Inoue M., Fujibayashi Y, & Saga T. (2015). Immuno-PET Imaging of HER3 in a Model in which HER3 Signaling Plays a Critical Role. PLoS ONE, 10(11), e0143076.

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Immunohistochemical Identification of Activated PSCs

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Formalin-fixed, paraffin embedded tumor tissues were sliced into 4-µm-thick sections and endogenous peroxidase activity was blocked with methanol containing 0.3% hydrogen peroxidase for 30 min at room temperature. Antigen retrieval was performed by boiling in a microwave oven (citrate buffer, pH 6.0). Afterwards, the tissues were incubated with mouse anti-α-smooth muscle actin (αSMA) antibody (1:100, #M0851; Dako) overnight at 4°C and stained with EnVision+System-HRP Labeled Polymer Anti-Mouse (#K4001; Dako). Activated PSCs were identified based on their spindle-like shape and αSMA-positive staining. The αSMA-positive area was measured using analysis application Hybrid cell count (BZ-H3C, Keyence).

Matsumoto S., Nakata K., Sagara A., Guan W., Ikenaga N., Ohuchida K, & Nakamura M. (2021). Efficient pre-treatment for pancreatic cancer using chloroquine-loaded nanoparticles targeting pancreatic stellate cells. Oncology Letters, 22(2), 633.

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Immunohistochemical Evaluation of Claudin-7

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Four-micron sections from formalin-fixed, paraffin-embedded (FFPE) tissues were stained for claudin-7 after antigen retrival by microwave treatment in EDTA buffer-pH 8.0, applying anti-Claudin-7 (mouse, clone 5D10F3, 1:80, Thermo Scientific) for 60 min. The reaction was revealed using Novolink Polymer (Leica Microsystem) or Envision System-HRP Labeled Polymer anti-Mouse (Dako) followed by DAB.
Immunoreactivity was evaluated by four independent observers (CR, LA, MB, EB); staining was graded for intensity (1–3+) and percentage of stained cells, and a single H score was obtained by multiplying the intensity and the percentage staining. Positivity was defined by at least 1+ intensity in 10% or more of the cells (H-score = 10).

Romani C., Zizioli V., Silvestri M., Ardighieri L., Bugatti M., Corsini M., Todeschini P., Marchini S., D'Incalci M., Zanotti L., Ravaggi A., Facchetti F., Gambino A., Odicino F., Sartori E., Santin A.D., Mitola S., Bignotti E, & Calza S. (2020). Low Expression of Claudin-7 as Potential Predictor of Distant Metastases in High-Grade Serous Ovarian Carcinoma Patients. Frontiers in Oncology, 10, 1287.

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Immunohistochemical Analysis of Thyroid Tissue

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The resected thyroid glands were fixed in 20% buffered formalin and then embedded in paraffin blocks to cut sections for immunohistochemistry. Deparaffinized 4-μm sections were pre-treated by autoclaving for 20 min at 120°C in target retrieval solution (pH 6.0) (Dako, Glostrup, Denmark). The sections were incubated for 1 h at 37°C with 1:50 dilution of anti-Ki-67 mouse monoclonal antibody (MIB-5, Dako) or 1:200 dilution of anti-p21 mouse monoclonal antibody (CP74, Abcam, Cambridge, UK) in a humidified chamber. After incubation with EnVision+ system-HRP-labeled polymer anti-mouse (Dako) for 60 min, immunoreactivity was visualized by incubation with 3,3-diaminobenzidine (DAB) using a liquid DAB+ substrate chromogen system (Dako). The number of Ki-67-positive cells in thyroid follicular cells was counted as the Ki-67 labeling index (LI; the number of positive cells per one high-power field). The level of p21 immunoreactivity in thyroid follicular cells was assessed by scoring the highest intensity of staining in one high-power field as follows: negative, 0; faint nuclear staining, 1; moderate nuclear staining, 2; intense nuclear and cytoplasmic staining, 3.

Kurohama H., Matsuda K., Kishino M., Yoshino M., Yamaguchi Y., Matsuu-Matsuyama M., Kondo H., Mitsutake N., Kinoshita A., Yoshiura K.I, & Nakashima M. (2021). Comprehensive analysis for detecting radiation-specific molecules expressed during radiation-induced rat thyroid carcinogenesis. Journal of Radiation Research, 62(Suppl 1), i78-i87.

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HER2 IHC Evaluation Protocol

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The primary antibodies used to detect the cytoplasmic domain of HER2 were purchased from Merck and Abcam, and the antibody used to detect the external domain of HER2 was purchased from Abnova. All of the collected tissues were fixed in FFPE blocks. Xenograft and cell-block sections were cut at 3 μm and human sections were cut at 4 μm for the HER2 IHC study. Paraffin sections were dewaxed and rehydrated in a Leica XL autostainer. Following antigen retrieval, the sections were incubated with 10 min of endogenous peroxidase block (DAKO), 60 min of primary antibodies, 30 min of EnVision System-HRP labeled polymer anti-mouse (DAKO), and 10 min of diaminobenzidine substrate (DAKO K3468), in that order. Finally, the sections were counter-stained, dehydrated, cleared, and mounted with coverslips in a Leica XL autostainer workstation. A HercepTest™ (DAKO) was used to detect the membranous expression of HER2, following standard procedures. Each slide was evaluated and scored on a 0–3 scale, following uniform guidelines developed for GC HER2 scoring from ToGA trials [13 (link)].

Yu D.H., Tang L., Dong H., Dong Z., Zhang L., Fu J., Su X., Zhang T., Fu H., Han L., Xie L., Chen H., Qian Z., Zhu G., Wang J., Ye Q., Zhang J., Yin X., Zhang X., Ji J, & Ji Q. (2015). Oncogenic HER2 fusions in gastric cancer. Journal of Translational Medicine, 13, 116.

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Immunohistochemical Analysis of GLP-1R in Pancreatic Cancer

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Pancreatic tissues and immunohistochemical staining. The study was approved by the Ethics Committee of Kyushu University (Fukuoka, Japan) and was conducted according to the Ethical Guidelines for Human Genome/Gene Research enacted by the Japanese Government and the Helsinki Declaration. Tissue samples of pancreatic cancer were obtained from 48 patients who underwent surgery for PDAC at Kyushu University Hospital between 2000 and 2009. Paraffin-embedded sections from the tissue samples were deparaffinized in xylene and rehydrated in a graded ethanol series. After endogenous peroxidase activity was blocked by incubation with 3% hydrogen peroxidase in methanol for 30 min, antigen retrieval was achieved by microwaving the sections in citrate buffer for 20 min, and the samples were then incubated overnight at 4˚C with a mouse monoclonal anti-human GLP-1R antibody (clone 419208; dilution, 1:100; R&D Systems, Minneapolis, MN, USA). For immunohistochemical labeling, we employed a peroxidase-labeled polymer (EnVision™+ System-HRP Labeled Polymer Anti-Mouse; Dako, Carpinteria, CA, USA) and used 3,3'-diaminobenzidine tetrahydrochloride (Sigma-Aldrich, St. Louis, MO, USA) as a chromogen. Finally, the sections were counterstained with hematoxylin. Negative control staining without the primary antibody was performed to confirm the antibody specificity.

Cases A.I., Ohtsuka T., Kimura H., Zheng B., Shindo K., Oda Y., Mizumoto K., Nakamura M, & Tanaka M. (2015). Significance of expression of glucagon-like peptide 1 receptor in pancreatic cancer. Oncology reports, 34(4).

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Immunohistochemical Analysis of DSSP Expression

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Specimens were washed with DPBS and fixed in 4% buffered formaldehyde solution for 3 hours at 4°C. Decalcification of scaffolds was performed by incubation in a solution containing 90% Osteosoft (Merck KGaA, Darmstadt, Germany) and 10% of the above formaldehyde solution for up to 21 days. Samples were then embedded in paraffin, cut into sections, and subjected to hematoxylin and eosin and Masson’s trichrome staining according to standard protocols.
DSSP expression was evaluated by immunohistochemistry using a mouse anti-human antibody (LFMb-21, dilution 1:50; Santa Cruz Biotechnology, Santa Cruz, CA) according to the manufacturer’s instructions. Sections were deparaffinized and rehydrated through a graded ethanol series, rinsed in distilled water, and treated with 0.3% H₂O₂ and 10% normal horse serum to block endogenous peroxidase and nonspecific binding, respectively. The Envision amplification system (Envision System + labeled polymer-HRP anti-mouse; Dako, Carpinteria, CA, USA) was used, followed by development with 3,3′-diaminobenzidine (Dako) as a chromogen according to the manufacturer’s instructions, which yielded brown staining in immunoreactive structures. Sections were finally counterstained with Mayer’s hematoxylin (Sigma-Aldrich).

Martín-de-Llano J.J., Mata M., Peydró S., Peydró A, & Carda C. (2019). Dentin tubule orientation determines odontoblastic differentiation in vitro: A morphological study. PLoS ONE, 14(5), e0215780.

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Immunohistochemical Analysis of DSSP Expression

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Specimens were washed with DPBS and fixed in 4% buffered formaldehyde solution for 3 h at 4 °C. Decalcification of the scaffolds was performed by incubation in a solution containing 90% Osteosoft (Merck KGaA, Darmstadt, Germany) and 10% formaldehyde solution for up to 21 days. Samples were then embedded in paraffin, cut into 5-µm sections, and stained with hematoxylin and eosin and Masson’s trichrome according to standard protocols.
DSSP expression was evaluated by immunohistochemistry using a mouse anti-human antibody (LFMb-21, dilution 1:50; Santa Cruz Biotechnology, Santa Cruz, CA, USA) according to the manufacturer’s instructions. Sections were deparaffinized and rehydrated using a graded ethanol series, rinsed with distilled water, and treated successively with 0.3% H₂O₂ and 10% normal horse serum to block endogenous peroxidase and nonspecific binding, respectively. The Envision amplification system (Envision System + labeled polymer-HRP anti-mouse; Dako, Carpinteria, CA, USA) was used, followed by development with 3,3′-diaminobenzidine (Dako) as chromogen according to the manufacturer’s instructions, which produced brown staining in immunoreactive structures. Sections were finally counterstained with Mayer’s hematoxylin (Sigma-Aldrich).

Mata M., Peydró S., de Llano J.J., Sancho-Tello M, & Carda C. (2022). Human Dental Pulp Stem Cells Differentiate into Cementoid-Like-Secreting Cells on Decellularized Teeth Scaffolds. International Journal of Molecular Sciences, 23(24), 15588.

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