Endogenous peroxidase block

Manufactured by Agilent Technologies

Sourced in United Kingdom

Endogenous peroxidase block is a laboratory reagent used to inhibit the activity of endogenous peroxidase enzymes in biological samples prior to immunohistochemical or other enzymatic staining procedures. This aids in reducing background staining and improving the specificity of the target signal detection.

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Lab products found in correlation

Autostainer xl, by Leica (1 mentions) Envision system hrp labeled polymer anti mouse, by Agilent Technologies (1 mentions) Herceptest, by Agilent Technologies (1 mentions) Sc 52746, by Santa Cruz Biotechnology (1 mentions) Dako autostainer, by Agilent Technologies (1 mentions) Novared chromagen, by Vector Laboratories (1 mentions) Envision flex wash buffer, by Agilent Technologies (1 mentions) Biotinylated goat anti mouse igg, by Vector Laboratories (1 mentions) Cytoseal xyl, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Endogenous peroxidase Peroxidase block

3 protocols using endogenous peroxidase block

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 Staining of TNFα and Integrin β7

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Tissues were deparaffinized and rehydrated to water, and after a low-temperature retrieval technique, ALTER,^{24 (link)} they were immunostained on a Dako Autostainer (Dako, Stockport, UK). Briefly, staining comprised a 10-minute endogenous peroxidase block (Dako) followed by a 10-minute protein block in 2% casein (Vector Labs, Birmingham, UK). Sections were incubated in optimally diluted antibodies for 1 hour; mouse anti-TNFα, 1/200 (sc52746, Santa Cruz Biotechnology, CA, USA) and rabbit anti-integrin b7, 1/400 (HPA042277; Sigma-Aldrich, Gillingham, UK). Antibody detection was performed with Vector Excel mouse and rabbit kits, respectively and visualized in NovaRED chromagen (Vector Labs) for 5 minutes. All buffer washes performed were with EnVision FLEX wash buffer (Dako). Sections were then counterstained with Meyers hematoxylin, dehydrated through to xylene, and mounted with a glass coverslip in distyrene plasticizer xylene. Expression was scored as the following: 0 = none, 1 = low (<30%), 2 = medium (30%-60%), and 3 = high (>60%).

Iacucci M., Jeffery L., Acharjee A., Grisan E., Buda A., Nardone O.M., Smith S.C., Labarile N., Zardo D., Ungar B., Hunter S., Mao R., Cannatelli R., Shivaji U.N., Parigi T.L., Reynolds G.M., Gkoutos G.V, & Ghosh S. (2022). Computer-Aided Imaging Analysis of Probe-Based Confocal Laser Endomicroscopy With Molecular Labeling and Gene Expression Identifies Markers of Response to Biological Therapy in IBD Patients: The Endo-Omics Study. Inflammatory Bowel Diseases, 29(9), 1409-1420.

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Immunohistochemical Staining of GD2 Antigen

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Frozen tissue samples were obtained from surgical samples at the time of clinically indicated tumor resection or biopsy after informed consent. Sections were sliced at 5-micron thickness, fixed in cold acetone for 10 minutes, dried at room temperature for 5 minutes, and rehydrated in PBS for 10 minutes. Samples were quenched with endogenous peroxidase block (DAKO) for 10 minutes, washed for 5 minutes, and incubated for 60 minutes at 37°C with mouse anti-human GD2 (14G2a, Millipore) diluted in DAKO’s antibody diluent with background reducing components to a 1:100 concentration. After washing, sections were incubated for 30 minutes in anti-mouse (biotinylated goat anti-mouse IgG, Vector, Burlingame, CA) at a concentration of 2.5 μg/mL. Sections were washed with DAKO wash buffer, incubated in DAKO peroxidase substrate solution for 5 minutes, and washed in PBS. The reaction was developed by a 2-5 minute incubation with the of 3’-3’ diaminobenzidine chromogenic solution (Vector). Slides were then counterstained with hematoxylin, dehydrated with a series of alcohol solutions (50%-100%), followed by three changes of xylene and mounted with Cytoseal XYL (Thermo Scientific). Analysis was performed using standard microscopy.

Long A.H., Highfill S.L., Cui Y., Smith J.P., Walker A.J., Ramakrishna S., El-Etriby R., Galli S., Tsokos M.G., Orentas R.J, & Mackall C.L. (2016). Reduction of MDSCs with all-trans retinoic acid improves CAR therapy efficacy for sarcomas. Cancer immunology research, 4(10), 869-880.

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