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Endogenous peroxidase blocking reagent

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The Endogenous peroxidase-blocking reagent is a laboratory product designed to inhibit endogenous peroxidase activity in tissue sections or cell preparations prior to immunohistochemical staining procedures. This reagent helps to eliminate the interference of endogenous peroxidases, which can lead to non-specific staining and false-positive results.

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Lab products found in correlation

Lsm 780, by Zeiss (4 mentions) Protein block, by Agilent Technologies (2 mentions) Antigen retrieval solution, by Agilent Technologies (1 mentions) Alexa fluor 488 conjugate, by Thermo Fisher Scientific (1 mentions) Vectashield h 1200, by Vector Laboratories (1 mentions) Proteinase k, by Agilent Technologies (1 mentions)

Close spelling variants of this product

Peroxidase blocking reagent (80 citations) Peroxidase blocking (19 citations) Blocking reagent (15 citations) Endogenous peroxidase blocking (12 citations) Endogenous peroxidase (8 citations)

4 protocols using endogenous peroxidase blocking reagent

1

Histological and Immunohistochemical Analysis of Human Tissues

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For histological and immunohistochemical analysis, human tissues were fixed with 4% PFA and embedded in paraffin. 4 μm sections were deparaffinized and antigens retrieved with 10mM sodium citrate. Sections were treated with endogenous peroxidase-blocking reagent (Dako) for 5 minutes and protein block (Dako) for 10 minutes at room temperature. Sections were incubated with DLX3 antibody (Abcam ab178428, 1:500 dilution) overnight at 4°C, followed by incubation with horseradish peroxidase-labeled secondary Abs (ENVISION: Dako) and visualized with 3, 3′-diaminobenzidine tetra-hydrochloride, counterstained with Mayer’s hematoxylin.
For immunofluorescence staining, sections were blocked with 3% dry milk-PBS supplemented with 5% normal goat serum for 1 hour at room temperature. Antibody details are in Supplementary Table 1. Sections were counterstained with 4,6-diamidino-2-phenylindole (DAPI) to visualize nuclei.
Proliferative activity in tumors was assessed by calculating the percentage of PCNA-positive nuclei examined using laser-scanning confocal microscope Zeiss LSM780. The number of positive cells was expressed as a percentage of the total number counted to give a PCNA score. Two independent sections per animal were stained and analyzed. A minimum of five microscopic fields per section were counted manually by two independent investigators and the mean was used for analysis.
Bajpai D., Mehdizadeh S., Uchiyama A., Inoue Y., Sawaya A., Overmiller A., Brooks S.R., Hasneen K., Kellett M., Palazzo E., Motegi S.I., Yuspa S.H., Cataisson C, & Morasso M.I. (2021). Loss of DLX3 tumor suppressive function promotes progression of SCC through EGFR-ERBB2 pathway. Oncogene, 40(21), 3680-3694.
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2

Immunohistochemical Analysis of KOPr in Brain Regions

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The animals were killed by cervical dislocation immediately after the CPP experiment was completed. The brain tissues were dissected out, which was followed by the tissue processing.[2 ] Only four parts of brain regions were used during immunohistochemistry (IHC), namely amygdala, hippocampus, prefrontal cortex, and striatum. Tissue sections were incubated with antigen retrieval solution (Dako, Santa Clara, CA, USA) using a commercial microwave for 10 min. Then, the tissues were incubated with ready-to-use endogenous peroxidase blocking reagent (Dako) for 10 min at room temperature. Next, the tissues were incubated with primary antibody (rabbit monoclonal antibody [EPR18881] to KOPr) with dilution factor of 1:500 for 30 min. The slides were then incubated with Dako Real EnVision HRP (horse radish peroxidase) polymer (ready-to-use secondary antibody) for 30 min. The tissues were then incubated with 3,3’-diaminobenzidine (DAB) substrate (1mL DAB substrate + 20 μL DAB chromogen) for 10 min.
Wasli N.S., Ridzwan I.E., Azzubaidi M.S., Kasmuri A.R., Ahmed Q.U., Ming L.C., Mohamed N, & Syd Mohmad Faudzi S.M. (2020). Striatum Hyperactivity Triggers Relapse to Morphine and Methamphetamine (Polydrug) Dependence in Mice. Journal of Pharmacy & Bioallied Sciences, 12(Suppl 2), S826-S830.
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3

Histological and Immunohistochemical Analysis of Human Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols
For histological and immunohistochemical analysis, human tissues were fixed with 4% PFA and embedded in paraffin. 4 μm sections were deparaffinized and antigens retrieved with 10mM sodium citrate. Sections were treated with endogenous peroxidase-blocking reagent (Dako) for 5 minutes and protein block (Dako) for 10 minutes at room temperature. Sections were incubated with DLX3 antibody (Abcam ab178428, 1:500 dilution) overnight at 4°C, followed by incubation with horseradish peroxidase-labeled secondary Abs (ENVISION: Dako) and visualized with 3, 3′-diaminobenzidine tetra-hydrochloride, counterstained with Mayer’s hematoxylin.
For immunofluorescence staining, sections were blocked with 3% dry milk-PBS supplemented with 5% normal goat serum for 1 hour at room temperature. Antibody details are in Supplementary Table 1. Sections were counterstained with 4,6-diamidino-2-phenylindole (DAPI) to visualize nuclei.
Proliferative activity in tumors was assessed by calculating the percentage of PCNA-positive nuclei examined using laser-scanning confocal microscope Zeiss LSM780. The number of positive cells was expressed as a percentage of the total number counted to give a PCNA score. Two independent sections per animal were stained and analyzed. A minimum of five microscopic fields per section were counted manually by two independent investigators and the mean was used for analysis.
Bajpai D., Mehdizadeh S., Uchiyama A., Inoue Y., Sawaya A., Overmiller A., Brooks S.R., Hasneen K., Kellett M., Palazzo E., Motegi S.I., Yuspa S.H., Cataisson C, & Morasso M.I. (2021). Loss of DLX3 tumor suppressive function promotes progression of SCC through EGFR-ERBB2 pathway. Oncogene, 40(21), 3680-3694.
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4

Collagen and Stem Cell Analysis in Wounded Skin

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Paraffin-embedded tissues were cut into 5 μm sections, deparaffinized in xylene, and rehydrated in PBS. Deparaffinized sections were treated with endogenous peroxidase-blocking reagent (DAKO Cytomation A/S, Copenhagen, Denmark) and proteinase K (DAKO) for 6 min at room temperature. Sections were then incubated with rabbit monoclonal antibody (mAb) specific for Collagen-1 (LSL, Tokyo, Japan), Collagen-3 (LSL) or Collagen-5 (BIOSS, Woburn, MA, USA), p75NTR/CD271 (Chemicon International, Carlsbad, CA, USA), and DAPI (VECTASHIELD H-1200; Vector Laboratories, Burlingame, CA, USA) was used for nuclear staining. In some experiments, non-wounded and wounded skin sections were double-stained using a mAb specific for CD45 (Merck Millipore Corporation, Darmstadt, Germany) and rabbit mAb specific for CD271. Sections were sequentially incubated with goat anti-Rabbit IgG (H+L) or goat anti-Mouse IgG (H+L) secondary Ab, Alexa Fluor ® 488 conjugate (Invitrogen Corp., CA, USA). Sections were washed 3 times with PBS between incubations. For the analysis of accumulation of collagen-1, -3, and -5, fluorescence intensity/100 µm 2 of dermis was measured using a NIH-image software (USA National Institutes of Health). For analysis of dermal CD271 + cells, stained cells were counted in 200 µm 2 of the wound bed per section.
Iwata Y., Hasebe Y., Hasegawa S., Nakata S., Yagami A., Matsunaga K., Sugiura K, & Akamatsu H. (2017). Dermal CD271+ Cells are Closely Associated with Regeneration of the Dermis in the Wound Healing Process. Acta dermato-venereologica, 97(5).
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