The following products were from Sigma Aldrich unless otherwise stated: Thymidine (T1895), BrdU (10 mg/ml stock in H2O, freshly prepared, used at 30 μM; B5002), Hydroxyurea (HU, H8627), H2SO4 (320501), Butyric acid (1 M stock in DMSO; B5887), Trichostatin (TSA, 2 mg/ml – 6.6 mM stock in DMSO; T8552), Bleomycin (10 mg/ml stock in DMSO; B2434), Propionic anhydride (81942), Lactic acid, acetonitrile, ammonium bicarbonate, ICRF (2 mg/ml stock in DMSO, used at 2 μg/ml), Camptothecin (5 mM stock in DMSO, 1100/25, R&D Systems Europe), Etoposide (85 mM in DMSO stock; E1383), gelatin (G9391), FLAG immunoprecipitation kit (FLAGIPT1-1KT), acetyl-CoA (A2056-1mg). Micrococcal nuclease S7 (10107921001; Roche), stock 15,000 U ml−1 in MNase buffer (see below). Chk1 inhibitors: AZD7762 (used at 50 or 500 nM), CHIR-124 (15 mM stock in DMSO, used at 1 μM), and SCH900776 (150 mM stock in DMSO, used at 10 μM) were from Seleckchem Euromedex (S1532, S2683 and S2735, respectively). Trypsin Gold MS grade (V5280; Promega), pyridostatin pentahydrochloride (4763; R&D Systems, used at 10 μM), recombinant full length H3 (C110A, 31207; Active Motif); highly active recombinant His-tagged CREB-binding Protein (mouse; CPB, CREBBP) was from AdipoGen (AG-40T-0016; A00269/C).
S7 micrococcal nuclease
Manufactured by Roche
The S7 Micrococcal nuclease is a laboratory equipment product designed for specific applications. It functions as an enzyme that cleaves DNA and RNA, facilitating various research and analysis processes. The core function of this product is to provide a reliable tool for nucleic acid manipulation and processing, supporting a range of scientific endeavors.
Automatically generated - may contain errors
Lab products found in correlation
Mirvana mirna isolation kit, by Thermo Fisher Scientific (2 mentions)
Bafilomycin a1, by Merck Group (2 mentions)
Ly294002, by Merck Group (2 mentions)
Human m csf, by Thermo Fisher Scientific (2 mentions)
Cytometric bead array cba human inflammatory cytokine kit, by BD (2 mentions)
Human ifn γ, by Roche (2 mentions)
Ultrapure e coli lps, by InvivoGen (2 mentions)
Torin1, by R&D Systems (2 mentions)
Myc sc 764, by Santa Cruz Biotechnology (2 mentions)
Leucyl trna synthetase ab31534, by Abcam (2 mentions)
P tsc2 thr1462, by Cell Signaling Technology (2 mentions)
Complete protease inhibitor cocktail, by Roche (2 mentions)
Trypsin gold ms grade, by Promega (1 mentions)
Thymidine, by Merck Group (1 mentions)
Bleomycin, by R&D Systems (1 mentions)
H2so4, by R&D Systems (1 mentions)
Etoposide, by R&D Systems (1 mentions)
Pierce anti ha magnetic beads, by Thermo Fisher Scientific (1 mentions)
Bioruptor ultrasonic cell disruptor, by Diagenode (1 mentions)
9 protocols using s7 micrococcal nuclease
1
Comprehensive Cell Cycle Regulation Assay
2
Neutrophil Extracellular Trap Formation
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Neutrophil extracellular traps (NETs) were determined on 0.25×106 PMN. Cells were treated or untreated with Stx2 and/or LPS, or in the presence of non-stimulated (washed) or treated Plts as described above (PMN/Plts ratio, 1:50). Treated PMNs were left for 6 h, and NETs were measured using different approaches as detailed below. Micrococcal Nuclease S7 (4 U, Hoffmann-La Roche, Basilea, Suiza) was used as a control to digest NETs.
3
Micrococcal Nuclease S7 Administration in Mice
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Control and LPS/Stx2-treated mice received an i.v. injection of Micrococcal Nuclease S7 (Hoffmann-La Roche, Basilea, Suiza) (150 U in buffer containing 5 nM Ca++ and 5 mM Mg++) every 12 h. The first dose was given 1 h before LPS/Stx2 treatment.
4
Multi-omics Profiling of Cell Signaling
5
Multi-omics Profiling of Cell Signaling
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Human IFN-γ and S7 micrococcal nuclease were purchased from Roche; human M-CSF was from Peprotech; Pam3CSK4 was from EMC Microcollections; UltraPure E.Coli LPS was from Invivogen. MG132, CGP57380, Rapamycin and PP242 were purchased from Calbiochem; Cycloheximide, Bafilomycin A1, 1-Methyl-D-tryptophan (1-D-MT), L-Tryptophan and 10058-F4 were from Sigma; Torin 1 was from R&D Systems; LY 294002 was from EMD Millipore. The following antibodies were purchased from Cell Signaling: mTOR (#2983), p-TSC2(Thr1462) (#3617), TSC2 (#4308), p-Akt (Ser473) (#4060), p-p70S6K(Thr421/Ser424) (#9204), 4E-BP1 (#9644), p-4E-BP1(Thr37/46) (#2855), LC3A (#4599), LC3B (#3868), IκB-α (#9242), Mnk1(#2195), p-Mnk1(Thr197/202) (#2111), eIF4E (#9742), p-eIF4E(Ser209) (#9741), p-p38(Thr180/Tyr182) (#9215), p-ERK1/2 (Thr202/Tyr204) (#9101), ERK (#9102), β-catenin (#9562), M-CSFR (#3152). Additionally, p38α (sc-535), HES1 (sc-25392), TBP (sc-204), LAMP1 (sc-20011) and Myc (sc-764) were from Santa Cruz Biotech. PABPC1 (ab6125), β-tubulin (ab11307) and Leucyl tRNA synthetase (ab31534) antibodies were from Abcam. mirVana miRNA isolation kit was purchased from Ambion/Life Technologies. BD™ Cytometric Bead Array (CBA) human inflammatory cytokine kit was purchased from BD Biosciences.
6
Chromatin Fractionation and MNase Digestion of Embryonic Stem Cells
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Embryonic stem cells were collected by trypsinization, washed once with PBS and counted. ES cell pellets were resuspended at a concentration of 10mio cells/ml in chromatin fractionation buffer (10 mM Hepes pH 7.6, 150 mM NaCl, 3 mM MgCl2, 0.5% Triton X‐100, 1 mM DTT freshly supplemented with cOmplete™ Protease Inhibitor Cocktail (Roche)) and incubated for 30 min at room temperature rotating. Precipitated chromatin was fractionated by centrifugation. Total and chromatin fractionated samples were further processed by MNase (S7 Micrococcal nuclease, Roche) digest for ensuring sufficient genomic DNA fragmentation. All samples were incubated in 1× Laemmli buffer (10% glycerol, 10 mM Tris pH 6.8, 2% SDS, 0.1 mg/ml bromphenolblue, 2% β‐mercaptoethanol) at 95°C for 5 min and were further analyzed by Western Blotting.
7
Chromatin Fractionation and Western Blot
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Cells were trypsinized, resuspended in the chromatin fractionation buffer (10 mM Hepes [pH = 7.6], 150 mM NaCl, 3 mM MgCl2, 0.5%Triton X-100, 1 mM dithiothreitol supplemented with cOmplete™ proteinase inhibitor compound) (Roche, CA, USA) at 1.0 × 107 cells/mL and incubated for 30 min. Precipitated chromatin was centrifuged, and total and chromatin fractionated samples were treated with MNase (S7 Micrococcal nuclease, Roche). All samples were incubated in 1 × Laemmli buffer (10% glycerol, 10 mM Tris [pH = 6.8], 2% sodium dodecyl sulfate [SDS], 0.1 mg/ml bromphenolblue, 2% β-mercaptoethanol) at 95 °C for 5 min and were further analyzed using Western Blot Dalcher et al. (2020 (link)).
8
Chromatin Immunoprecipitation (ChIP) Protocol
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ChIP experiments were performed as previously described (Leone et al, 2017 (link); Peña-Hernández et al, 2021 (link)). Briefly, 1% formaldehyde was added to cultured cells to cross-link proteins to DNA. Isolated nuclei were then digested with MNase (S7 Micrococcal nuclease; Roche) and briefly sonicated using a Bioruptor ultrasonic cell disruptor (Diagenode) to shear genomic DNA to an average fragment size of 200 bp. 200 μg of chromatin was diluted tenfold with ChIP buffer (16.7 mM Tris–HCl pH 8.1, 167 mM NaCl, 1.2 mM EDTA, 0.01% SDS, and 1.1% Triton X-100) and precleared for 2 h with 10 μl of packed Sepharose beads for at least 2 h at 4°C. Immunoprecipitation was performed overnight with the HA-magnetic beads (Pierce Anti-HA magnetic beads; Thermo Fisher Scientific). After washing, elution, and reversion of cross-links, the eluates were treated with RNase A (1 μg). DNA was purified with phenol-chloroform, ethanol precipitated, and quantified by quantitative PCR. Primer sequences and antibodies are listed in Table S10.
9
Chromatin Fractionation and MNase Digestion
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48 hours post-transfection, NIH3T3 cells were collected by trypsinization, washed once with PBS and counted. ES cell pellets were resuspended at a concentration of 10mio cells/ml in chromatin fractionation buffer (10mM Hepes pH 7.6, 150mM NaCl, 3mM MgCl2, 0.5% Triton X-100, 1mM DTT freshly supplemented with cOmplete ™ Protease Inhibitor Cocktail (Roche)) and incubated for 30 minutes at room temperature rotating. Precipitated chromatin was fractionated by centrifugation. Total and chromatin fractionated samples were further processed by MNase (S7 Micrococcal nuclease, Roche) digest for ensuring sufficient genomic DNA fragmentation. All samples were incubated in 1x Laemmli buffer (10% glycerol, 10mM Tris pH 6.8, 2% SDS, 0.1mg/ml bromophenolblue, 2% β-mercaptoethanol) at 95°C for 5 minutes and were further analyzed by Western Blotting.
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