Immunohistochemical staining with monoclonal mouse anti-human mitochondria (hMIT) (clone 113-1, Merck Millipore, Burlington, MA, 1:2000), antihuman CYP1A2 (clone 3B8C1, Abcam plc., Cambridge, UK, 1:1000), antihuman CYP2C9 (clone 2C8, LifeSpan Biosciences, Inc., Seattle WA. USA, 1:150), and rabbit antihuman CYP3A4 (clone EPR6202, Abcam Plc., 1:300) antibodies was performed as described previously14 (link). Tissues were fixed in 4% (v/v) phosphate-buffered formalin (Mildform 10 NM; Wako Pure Chemical Industries). The sections were autoclaved for 10 min in a target retrieval solution (0.1 M citrate buffer, pH 6.0; 1 mM EDTA, pH 9.0), equilibrated at room temperature for 20 min, and then incubated with an anti-POR (HPA010136, Sigma) primary antibody. Primary antibodies were visualized using amino acid polymer/peroxidase complex-labeled antibodies (Histofine Simple Stain MAX PO [MULTI]; Nichirei Biosciences Inc.) and diaminobenzidine (DAB; Dojindo Laboratories, Kumamoto, Japan) substrate (0.2 mg/mL 3,3-diaminobenzidine tetrahydrochloride in 0.05 M Tris–HCl, pH 7.6, and 0.005% H2O2). The sections were counterstained with hematoxylin. Images were captured using a digital slide scanner (NanoZoomer S60; Hamamatsu Photonics, KK Hamamatsu, Japan).
Mildform 10nm
Manufactured by Fujifilm
Sourced in Japan
The Mildform 10NM is a lab equipment product manufactured by Fujifilm. It is designed for precision measurement and analysis, but no further details about its core function can be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Histofine simple stain max po, by Nichirei Biosciences (1 mentions)
Hpa010136, by Merck Group (1 mentions)
Diaminobenzidine dab, by Dojindo Laboratories (1 mentions)
Nanozoomer s60, by Hamamatsu Photonics (1 mentions)
Accutase, by BD (1 mentions)
Click it edu imaging kit, by Thermo Fisher Scientific (1 mentions)
Dextran sulfate sodium (dss), by MP Biomedicals (1 mentions)
Midazolam, by Meiji Seika Pharma (1 mentions)
Butorphanol, by Meiji Seika Pharma (1 mentions)
Optimal cutting temperature medium, by Sakura Finetek (1 mentions)
4 protocols using mildform 10nm
1
Immunohistochemical Staining of Cytochrome P450 Enzymes
2
Teratoma Formation Assay for iPSCs
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The cultured cells were treated with Accutase, and feeder cells were removed by using a cell strainer (35 μm; Falcon, Corning, NY, USA). Approximately 1 × 106 cells were
resuspended in iPSC medium without inhibitors or pLIF. The medium was carefully removed after centrifuging. The cell pellet was xenografted into the testis of SCID mice using a fine glass
capillary with a mouthpiece. Three to five mice per clone were used, and teratoma formation was examined 12 weeks later. Mice were maintained and used in accordance with guidelines issued by
the NARO Institutional Animal Care and Use Committee. Teratomas were excised and fixed in Mildform 10NM (FUJIFILM Wako Pure Chemical, Osaka, Japan). A small piece of each teratoma was
embedded in paraffin, sectioned, stained using hematoxylin and eosin (HE), and examined using a microscope [23 (link)].
resuspended in iPSC medium without inhibitors or pLIF. The medium was carefully removed after centrifuging. The cell pellet was xenografted into the testis of SCID mice using a fine glass
capillary with a mouthpiece. Three to five mice per clone were used, and teratoma formation was examined 12 weeks later. Mice were maintained and used in accordance with guidelines issued by
the NARO Institutional Animal Care and Use Committee. Teratomas were excised and fixed in Mildform 10NM (FUJIFILM Wako Pure Chemical, Osaka, Japan). A small piece of each teratoma was
embedded in paraffin, sectioned, stained using hematoxylin and eosin (HE), and examined using a microscope [23 (link)].
3
Dextran Sulfate Sodium-Induced Colitis in Mice
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Mice received 1.25 or 0.75% DSS (molecular weight: 36,000-50,000, MP Biomedicals, Santa Ana, CA, USA) in drinking water for 7 days, followed by regular water for 5 days. Body weight, water consumption, and clinical scores were monitored daily. Clinical scores were determined as described for the C. rodentium infection model.
In vivo EdU incorporation assay WT and Spib -/-mice were killed 4 h or 2 days after intraperitoneal injection of 1 mg EdU in PBS, and colons were embedded in O.C.T. compound. Slices were fixed in Mildform 10NM (Wako) for 15 min and permeabilized with 0.5% Triton X-100 in PBS for 20 min. EdU was detected using Click-iT EdU Imaging Kits (Invitrogen) following the manufacturer's instructions.
In vivo EdU incorporation assay WT and Spib -/-mice were killed 4 h or 2 days after intraperitoneal injection of 1 mg EdU in PBS, and colons were embedded in O.C.T. compound. Slices were fixed in Mildform 10NM (Wako) for 15 min and permeabilized with 0.5% Triton X-100 in PBS for 20 min. EdU was detected using Click-iT EdU Imaging Kits (Invitrogen) following the manufacturer's instructions.
4
Euthanasia and Tissue Fixation of Common Marmosets
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Common marmosets were euthanized by exsanguination under deep anesthesia with a mixture of medetomizine (120 μg/kg), midazolam (600 μg/kg) and butorphanol (600 μg/kg; Meiji Seika Pharma Co., Ltd., Tokyo, Japan). Specimens of the GI tract were removed, washed with ice-cold PBS and fixed overnight with Mildform 10NM (Wako Pure Chemical Industries, Ltd., Osaka, Japan). Subsequently, tissue samples were soaked in 30% sucrose in PBS overnight at 40°C and frozen in Optimal Cutting Temperature medium (Sakura Finetek, Tokyo, Japan) at -80°C.
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