Dnase 1

Manufactured by Fujifilm

Sourced in Japan

DNase I is a laboratory enzyme used to degrade DNA. It functions by hydrolyzing the phosphodiester bonds in DNA, breaking down the DNA molecule into smaller fragments.

Automatically generated - may contain errors

Lab products found in correlation

Facs calibur hg flow cytometer, by BD (2 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (2 mentions) Cellquest software, by BD (2 mentions) Clone fjk 16s, by Abcam (2 mentions) Collagenase 1, by Fujifilm (2 mentions) Anti cd16 32 2.4g2, by BD (1 mentions) I a 1 e, by BD (1 mentions) Cd11b, by BioLegend (1 mentions) F4 80, by BioLegend (1 mentions) Cd11b pe antibody, by BD (1 mentions) Hyaluronidase, by Merck Group (1 mentions) Moflo astrio, by Beckman Coulter (1 mentions) Lysing buffer, by BD (1 mentions) Fc block, by BioLegend (1 mentions) Type 6 collagenase, by Merck Group (1 mentions) Streptavidin apc antibody, by Thermo Fisher Scientific (1 mentions) Iodoacetamide, by Fujifilm (1 mentions) Thiourea, by Fujifilm (1 mentions) Rnase a, by Fujifilm (1 mentions) Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions)

Close spelling variants of this product

DNase (7 citations) RNase-free DNase I (3 citations)

11 protocols using dnase 1

Phenotypic Analysis of Tumor Immune Cells

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Tumors were harvested and processed using Collagenase I and DNase I (Wako, Osaka, Japan). The resulting cell suspensions were clarified using 40-μm filters to prepare single cell suspensions, and single cells were suspended in PBS supplemented with 2% FBS. Splenocytes were hemolyzed and incubated with anti-CD16/32 2.4G2 (BD Biosciences, San Jose, CA, USA) to reduce FcγR binding. Cell-surface antigens were stained with antibodies specific for CD8 (BioLegend, San Diego, CA, USA, clone 53-6.7), CD4 (BioLegend, clone GK1.5), CD45 (BioLegend, clone 30-F11), IA/IE (BD Bioscience, clone M5/114), CD11c (BD Bioscience, clone HL3), CD11b (BioLegend, clone M1/70), and F4/80 (BioLegend, clone BM8).
For intracellular staining, cells were fixed and permeabilized using Foxp3/Transcription Factor Staining Buffer Set (eBioscience, San Diego, CA, USA) after cell surface staining and were stained with labeled antibodies against the intracellular molecules foxp3 (eBioscience, clone FJK-16s), glucose transporter 1 (GLUT1, Abcam, Cambridge, UK, clone EPR3915), and hexokinase II (HX2, Abcam, clone EPR20839).
Samples were analyzed on a FACS Calibur HG flow cytometer (BD Biosciences). Data analysis was performed with CellQuest™ software (Becton Dickinson, Lincoln Park, NJ, USA).

Tomita M., Yasui H., Higashikawa K., Nakajima K., Takakura H., Shiga T., Kuge Y, & Ogawa M. (2018). Anti PD-1 treatment increases [18F]FDG uptake by cancer cells in a mouse B16F10 melanoma model. EJNMMI Research, 8, 82.

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Quantifying Tumor-Infiltrating Immune Cells

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Mouse whole blood was collected with 1.5% EDTA-2Na used as the anticoagulation agent, and red blood cells were eliminated by a lysing buffer (BD Pharmingen). Leukocyte cells and BMDCs of normal control mice or OSC-19 tumour-bearing mice were stained with CD11b-PE antibody (BD Pharmingen). For IL-13Rα2 and CD11b double staining, OSC-19 tumours grown at non-IR sites or pre-IR sites in nude mice were minced and enzymatically digested with 3 mg/ml Type VI collagenase and 2 U/ml hyaluronidase (SIGMA). They were further treated with 0.1 mg/ml DNase I (Wako) and 0.09% NH₄Cl and strained through a 40 μm cell strainer. Single cells were incubated with IL-13Rα2-biotin (R&D systems) and CD11b-PE antibodies (BD Pharmingen), followed by Streptavidin-APC antibody (eBioscience). All samples were pre-incubated with Fc-block (BioLegend) prior to staining. Analysis was performed using MoFlo® Astrios (Beckman Coulter) and Summit Software ver. 4.3 (Beckman Coulter).

Okubo M., Kioi M., Nakashima H., Sugiura K., Mitsudo K., Aoki I., Taniguchi H, & Tohnai I. (2016). M2-polarized macrophages contribute to neovasculogenesis, leading to relapse of oral cancer following radiation. Scientific Reports, 6, 27548.

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Streptozotocin-based Diabetic Model

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Streptozotocin (STZ), urea, thiourea, sodium dodecyl sulfate (SDS), 3-((3-cholamidopropyl) dimethylammonio)-1-propanesulfonate (CHAPS), 2-mercaptoethanol (2-ME), dithiothreitol (DTT), bromophenol blue, iodoacetamide, RNase A, and DNase I were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Source information for all other assay reagents and materials is stated in the Materials and Methods section described below.

Matsuura K., Otani M., Takano M., Kadoyama K, & Matsuyama S. (2018). Proteomic Analysis of Hippocampus and Cortex in Streptozotocin-Induced Diabetic Model Mice Showing Dementia. Journal of Diabetes Research, 2018, 8953015.

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Immunofluorescence Staining of Rabbit Neutrophils

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Rabbit anti-human Rab27a polyclonal antibody (polyAb), mouse anti-α tubulin monoclonal antibody (mAb) was purchased from Sigma (St. Louis, MO). Mouse anti-human CD11b (complement receptor 3, CR3) mAb for flow cytometry was purchased from DAKO (Glostrup, Denmark). Rabbit anti-human MPO polyAb, rabbit anti-human histone H3 mAb (D1H2) and mouse anti-human histone H4 mAb (L64C1) was from Cell Signaling Technology (Danvers, MA). Rabbit anti-human histone H4 (citrulline 3) polyAb was from Millipore (Billerica, MA). Texas Red-conjugated zymosanA, 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H₂DCFDA) and Sytox Green were purchased from Lifetechnologies (Carlsbad, CA). Puromycin, phorbol 12-myristate 13-acetate(PMA) and diphenylene iodonium chloride (DPI), an NADPH oxidase inhibitor were purchased from Sigma (St. Louis, MO) and RPMI1640 medium was from Wako (Osaka, Japan). Penicillin-streptomycin mixed solution was from Nacalai Tesque (Kyoto, Japan), and polybrene was purchased from Millipore (Bedford, MA). Aminophenyl fluorescein (APF) and hydroxyphenyl fluorescein (HPF) were purchased from Sekisui Medical (Tokyo, Japan). Hoechst33342 was purchased from Dojindo Laboratories (Kumamoto, Japan). DNaseI was purchased from Wako (Osaka, Japan).

Kawakami T., He J., Morita H., Yokoyama K., Kaji H., Tanaka C., Suemori S.I., Tohyama K, & Tohyama Y. (2014). Rab27a Is Essential for the Formation of Neutrophil Extracellular Traps (NETs) in Neutrophil-Like Differentiated HL60 Cells. PLoS ONE, 9(1), e84704.

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Isolation of Mouse Testicular Cells

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Mouse testicular cells are obtained by the following method (La Salle et al., 2009) with some modifications. Briefly, a C57BL/6J mouse at 20 days postpartum (dpp) is euthanized using CO₂ gas, and testes are detunicated. The seminiferous tubules are placed in a 20 mL of KRB medium (120 mM NaCl, 4.8 mM KCl, 25.2 mM NaHCO₃, 1.2 mM KH₂PO₄, 1.2 mM MgSO₄•7H₂O, 1.3 mM CaCl₂, Pen/Strep (Gibco, 11548876), 11.1 mM D-glucose, essential amino acid (Gibco, 11130036), and nonessential amino acid (Gibco, 12084947), and containing collagenase type I (0.5 mg/mL, Wako, 035–17604), and incubated for 30 min at 32°C with shaking at 250 rpm. The seminiferous tubules are washed twice with a KRB medium. The tubules are further digested in 20 mL of fresh KRB medium containing Trypsin (0.5 mg/mL Wako, 207–19982) and DNase I (1 μg/mL, Wako, 314–08071) for 15 min at 32°C with shaking at 250 rpm to obtain a single-cell suspension. After a quick spin, cell pellets are washed twice with KRB medium containing 2% FBS and DNase I (1 μg/mL), and resuspended in 1× PBS (pH 6.8).

Fujiwara Y., Tanno Y., Sugishita H., Kishi Y., Makino Y, & Okada Y. (2021). Preparation of optimized concanavalin A-conjugated Dynabeads® magnetic beads for CUT&Tag. PLoS ONE, 16(11), e0259846.

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Astrocyte Culture from Neonatal Mouse Forebrain

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PCAs were prepared from neonatal C57BL/6J mice as described elsewhere [35 (link), 41 (link)]. Briefly, the isolated forebrain was minced and incubated with DNase I (FUJIFILM Wako Pure Chemical) and trypsin (Thermo Fisher Scientific) for dissociation. The dissociated tissues were suspended in Dulbecco’s modified Eagle medium/Nutrient Mixture F-12 (DMEM/F12; Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% penicillin/streptomycin (PS; Thermo Fisher). The suspended cells were seeded in poly-l-lysine (PLL; ScienCell, Carlsbad, CA, USA)-coated 75-mm² flasks (10–15 × 10⁶ cells/flask). The plated cells were cultured in a 5% CO₂ incubator at 37°C. Every 8–12 days, the confluent cells were shaken for 10 min to separate them from microglial cells, released from the flasks with trypsin, and seeded onto non-coated flasks. These cells were seeded onto non-coated 8-well chamber slides (2.5 × 10⁴ cells/well) and evaluated with immunocytochemistry for anti-glial fibrillary acidic protein (GFAP) antibody (Abcam).

Okazaki S., Boku S., Watanabe Y., Otsuka I., Horai T., Morikawa R., Kimura A., Shimmyo N., Tanifuji T., Someya T, & Hishimoto A. (2022). Polymorphisms in the hypoxia inducible factor binding site of the macrophage migration inhibitory factor gene promoter in schizophrenia. PLoS ONE, 17(3), e0265738.

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Immune Profiling of Tumor Microenvironment

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Thirteen mice were used for flow-cytometry (cGAMP alone group n = 6, cGAMP/anti-PD-1 combination group n = 7). Tumors were harvested and processed using Collagenase I and DNase I (Wako, Osaka, Japan). The resulting cell suspensions were clarified using 40-μm filters to prepare single cell suspensions, and single cells were suspended in PBS supplemented with 2% FBS. Splenocytes were hemolyzed and incubated with anti-CD16/32 2.4G2 antibody (BD Biosciences, San Jose, CA) to reduce FcγR binding. Cell-surface antigens were stained with antibodies specific for CD8 (BioLegend, San Diego, CA, clone 53-6.7), CD4 (BioLegend, clone GK1.5), and CD45 (BioLegend, clone 30-F11).
For intracellular staining, cells were fixed and permeabilized using a Foxp3/Transcription Factor Staining Buffer Set (eBioscience, San Diego, CA) after cell surface staining and then stained with labeled antibodies against the intracellular molecules Foxp3 (eBioscience, clone FJK-16s), glucose transporter 1 (GLUT1, Abcam, Cambridge, UK, clone EPR3915), and hexokinase II (HX2, Abcam, clone EPR20839).
Samples were analyzed on a FACS Calibur HG flow cytometer (BD Biosciences). Data analysis was performed with CellQuest™ software (Becton Dickinson, Lincoln Park, NJ).

Tomita M., Suzuki M., Kono Y., Nakajima K., Matsuda T., Kuge Y, & Ogawa M. (2020). Influence on [18F]FDG uptake by cancer cells after anti-PD-1 therapy in an enforced-immune activated mouse tumor. EJNMMI Research, 10, 24.

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Isolation and Purification of Total RNA from Tissue Specimens

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Total RNA from tissue specimens was prepared using ISOGEN (Nippon Gene, Tokyo, Japan), purified using RNeasy columns (Qiagen, Hilden, Germany) in accordance with manufacturer's instructions, and then treated with DNase I (Wako Pure Chemicals, Osaka, Japan). Total RNA was converted to cDNA by using random hexamers and SuperScript III reverse transcriptase (Invitrogen). The primer sets for detecting the expression of human UGT2 genes were described previously.¹⁵

Kobayashi K., Deguchi T., Abe S., Kajitani N., Kazuki K., Takehara S., Nakamura K., Kurihara A., Oshimura M, & Kazuki Y. (2022). Analysis of in vitro and in vivo metabolism of zidovudine and gemfibrozil in trans‐chromosomic mouse line expressing human UGT2 enzymes. Pharmacology Research & Perspectives, 10(6), e01030.

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Isolation of Intestinal Immune Cells

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Mice were euthanized by intraperitoneal injection of a large excess of pentobarbital sodium. Isolated colons were cut, opened longitudinally and washed with excess PBS to remove stools and mucus, followed by incubation for 30 min at 37°C with vigorous shaking in Ca²⁺- and Mg²⁺-free Hanks balanced salt solution supplemented with 5 mM EDTA and 1 mM DTT to remove epithelial layer. Cells released from colons were subjected to 40% / 70% Percoll gradient centrifugation and the cells containing IELs in middle layer were harvested and used for experiments. To isolate lamina propria cells, the remaining colons after removing epithelial layer were chopped into little pieces with scalpel and digested for 60 min at 37°C with gentle shaking in RPMI1640 medium supplemented with 5% FBS, 0.5 mg/ml Collagenase IV (SIGMA) and 50 U/ml DNase I (Wako). Cells were subjected to 40% / 70% Percoll gradient centrifugation and the cells in middle layer were harvested and used for experiments.

Sanjo H., Tokumaru S., Akira S, & Taki S. (2015). Conditional Deletion of TAK1 in T Cells Reveals a Pivotal Role of TCRαβ+ Intraepithelial Lymphocytes in Preventing Lymphopenia-Associated Colitis. PLoS ONE, 10(7), e0128761.

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Isolation and Culture of Pulmonary Endothelial Cells

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The PEA specimens and pulmonary arteries from lung lobectomy specimens were washed in sterile phosphate buffered saline (PBS), minced, and incubated in Dulbecco’s Modified Eagle Medium (DMEM) with 1% bovine serum albumin, 0.2% collagenase (Wako, Tokyo, Japan), 10 mg/ml DNaseI (Wako, Tokyo, Japan), 250 mg/ml dispase (Roche, Tokyo, Japan) for 1 h. The obtained cell suspensions were seeded into a 6-cm-petri dish coated with fibronectin (Corning, Corning, NY, USA) and incubated in an endothelial growth medium™-2 microvascular BulletKit™ (EGM; Lonza, Basel, Switzerland) until they reached approximately 80% confluence (7–10 days) at 37 °C in 5% CO₂ in an humidified air incubator. At the first passage, CD 31-positive ECs were isolated using CD31 MicroBeads (Miltenyi Biotec, Tokyo, Japan), as described previously [9 (link)]. The cells were incubated in EGM, and all of the experiments were carried out using cells that had been passaged less than 6 times.

Naito A., Sakao S., Lang I.M., Voelkel N.F., Jujo T., Ishida K., Sugiura T., Matsumiya G., Yoshino I., Tanabe N, & Tatsumi K. (2018). Endothelial cells from pulmonary endarterectomy specimens possess a high angiogenic potential and express high levels of hepatocyte growth factor. BMC Pulmonary Medicine, 18, 197.

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