Peroxidazed

Manufactured by Biocare Medical

Sourced in United States

Peroxidazed is a laboratory equipment that is used to detect the presence of peroxidase enzymes in biological samples. It is a versatile tool that can be used in various applications, such as immunohistochemistry, enzyme-linked immunosorbent assays (ELISA), and other peroxidase-based assays.

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Peroxidazed 1 (29 citations) Peroxidazed 1 (PX968; (5 citations) Peroxidazed 1 (PX968H, (2 citations)

4 protocols using peroxidazed

Histological and Immunohistochemical Analysis of Intestinal Tissue

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For routine histology, 4 μm sections of paraformaldehyde-fixed, paraffin-embedded, Swiss-rolled intestines were stained with haematoxylin and eosin. For immunohistochemical analyses, sections pretreated with citrate buffer at 95 °C, Peroxidazed (Biocare Medical, Pacheco, CA, USA), and Background Sniper (Biocare Medical, Pacheco, CA, USA) were incubated with antibodies recognizing β-catenin (BD, 610154), Ki67 (RTU, RM-9106-R7, Thermo Scientific) or pH3 (ser10) (ref. 9701S, Cell Signaling, Danvers, MA, USA), and then processed with the Vectastain Elite ABC Kit (PK6101) and the DAB Peroxidase Substrate Kit (SK4100, Vector Laboratories), or MACH 1 Universal HRP-Polymer Detection Kit and Betazoid DAB. The slides were stained with an intelliPATH flx automated staining system (Biocare Medical, Pacheco, CA, USA) and scanned with a MIRAX SCAN microscope with the MIRAX Control software (Zeiss, Oberkochen, Germany). Proliferating cells were quantified with the TissueMorph software and the TISSUEalign module (Visiopharm Integrator System version 5.0.2.1158, Hoersholm, Denmark) or the Analyze Particles function in ImageJ (version 1.8.0_172).

Zou Z.V., Le Gal K., El Zowalaty A.E., Pehlivanoglu L.E., Garellick V., Gul N., Ibrahim M.X., Bergh P.O., Henricsson M., Wiel C., Akyürek L.M., Bergo M.O., Sayin V.I, & Lindahl P. (2021). Antioxidants Promote Intestinal Tumor Progression in Mice. Antioxidants, 10(2), 241.

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Immunohistochemical Analysis of CXCL12/SDF-1 in FFPE Meninges

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Unstained formalin-fixed, paraffin-embedded (FFPE) human cerebral meningeal 5 μm sections were obtained from Amsbio. For antigen retrieval, slides were incubated in pH 6.0 buffer (Reveal Decloaking reagent, Biocare Medical) at 95–98°C for 30 minutes followed by a cool down period. Subsequent steps were then performed using an immunohistochemical staining platform (Intellipath, Biocare). Slides were immersed in a 3% hydrogen peroxide solution (Peroxidazed, Biocare) for 10 minutes, rinsed with TBST and blocked using a serum-free solution (Background Sniper, Biocare Medical, Concord, CA) for 10 minutes. The blocking solution was removed and the slides were incubated with human CXCL12/SDF-1 antibody (Clone D8G6H; Cell Signaling Technology) or IgG isotype control antibody (Cell Signaling Technology, #3900) 2.7 μg/mL in 10% blocking solution/90% TBST for 60 minutes at room temperature. Slides were then washed, and detection was performed using the Novocastra Novolink Polymer Kit (Leica Microsystems Inc.) according to the manufacturer’s specifications. Detection was performed with diaminobenzidine (DAB; Biolegend). Finally, the slides were counterstained with CAT hematoxylin (Biocare), dehydrated, and coverslipped.

Fujii N., Mushiake H, & Tanji J. (1998). An oculomotor representation area within the ventral premotor cortex. Proceedings of the National Academy of Sciences of the United States of America, 95(20), 12034-12037.

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Quantifying DKK1 Protein Levels in Cells

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DKK1 ELISAs were conducted using a Human DKK1 ELISA kit (Abcam: ab100501). Supernatants from each of the cell lines were analyzed in duplicate following the manufacturer’s protocol. Assay plates were read at 450 nm using the EnSpire Multimode Reader (PerkinElmer). A standard curve was generated using a four-parameter logistic regression fit. The amount of DKK1 was normalized to viable cell number which was determined prior to harvesting the supernatant. DKK1 IHC was performed at StageBio (Worcester, MA). FFPE CPA slide sections were subjected to high temperature antigen retrieval with DIVA Decloaker (Biocare Medical), followed by endogenous peroxidase blockade with Peroxidazed (Biocare Medical) and nonspecific protein binding blockade with Background Punisher (Biocare Medical). Subsequently, sections were incubated at room temperature for 2 h with an anti-DKK1 antibody (Cell Signaling: 4687) or a rabbit polyclonal IgG (Thermo-Fisher: 02-6102) at a concentration of 1.0 mg/mL. After washing, slides were incubated with a Rabbit-on-Farma HRP-Polymer (Biocare Medical) at room temperature for 30 min, washed, and developed for 5 min with a Betazoid DAB chromogen kit (Biocare Medical).

Caldwell C., Rottman J.B., Paces W., Bueche E., Reitsma S., Gibb J., Adisetiyo V., Haas M.S., Heath H., Newman W., Baum J., Gianani R, & Kagey M.H. (2021). Validation of a DKK1 RNAscope chromogenic in situ hybridization assay for gastric and gastroesophageal junction adenocarcinoma tumors. Scientific Reports, 11, 9920.

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Immunohistochemical Detection of CD99 in Human Meninges

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Unstained human meningeal sections (5 µm; Amsbio, T2234043) were de-paraffinized and rehydrate. For antigen retrieval, slides were incubated in pH 6.0 buffer (Reveal Decloaking reagent, Biocare Medical) at 95–98 °C followed for 30 min followed by a 20 min cool down period. Subsequent steps utilized an immunohistochemical staining platform (Intellipath, Biocare). Slides were immersed in a 3% hydrogen peroxide solution (Peroxidazed, Biocare) for 10 min, rinsed with TBST and blocked using a serum-free solution (Background Sniper, Biocare Medical, Concord, CA) for 10 min. Blocking solution was removed and slides were incubated with rabbit monoclonal anti-CD99 antibody (Clone EPR3097Y; Cell Marque) diluted in 10% blocking solution/90% TBST for 60 min at room temperature. Slides were then washed with TBST and detection performed with the Novocastra Novolink Polymer Kit (Leica Microsystems Inc.) according to the manufacturer’s specifications. Slides were then rinsed with TBST, detection performed with diaminobenzidine (DAB; Biolegend) and counterstained with CAT Hematoxylin (Biocare). Finally, slides were dehydrated and coverslipped.

Ebadi M., Jonart L.M., Ostergaard J, & Gordon P.M. (2021). CD99 antibody disrupts T-cell acute lymphoblastic leukemia adhesion to meningeal cells and attenuates chemoresistance. Scientific Reports, 11, 24374.

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