Thiazolyl blue tetrazolium bromide (MTT) and IGEPAL CA-630 were purchased from the USB Corporation (Cleveland, OH, USA). The medium, serum, and reagents for cell culture were obtained from Invitrogen Co. (Carlsbad, CA, USA). Other reagents used in this study were of reagent grade and were purchased from Sigma-Aldrich (St. Louis, MO, USA). K36H was supplied by one of the authors, Professor Kuo, and was synthesized as described previously [13 (link)].
Reagents for cell culture
Manufactured by Thermo Fisher Scientific
Sourced in United States, Poland
Reagents for cell culture are a range of chemical and biological components designed to support the growth and maintenance of cells in a laboratory environment. These reagents include media, supplements, and other essential elements required for cultivating cells in vitro. The core function of these reagents is to provide the necessary nutrients, growth factors, and environmental conditions to enable the proliferation and survival of cultured cells.
Automatically generated - may contain errors
Lab products found in correlation
Cell line, by American Type Culture Collection (3 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (3 mentions)
Pcdna3, by Promega (2 mentions)
Pcdna3.1 nfat2, by Promega (2 mentions)
Click it plus 5 ethynyl 2 deoxyuridine edu flow cytometry assay kit, by Thermo Fisher Scientific (2 mentions)
Pluronic f 127, by GeneCopoeia (2 mentions)
Hrp conjugated secondary antibody, by Bioss Antibodies (2 mentions)
C myc, by Cell Signaling Technology (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Fitc annexin 5 apoptosis detection kit, by BD (2 mentions)
Endotoxin free plasmid extraction kit, by Tiangen Biotech (2 mentions)
Roscovitine, by Merck Group (2 mentions)
Aav paov cmv bglobin vector 3flag, by Obio Technology (2 mentions)
Aav paov cmv bglobin cdt2 3flag, by Obio Technology (2 mentions)
Igepal ca 630, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Harvard Bioscience (1 mentions)
Cholesterol, by MP Biomedicals (1 mentions)
Plasticware, by Corning (1 mentions)
Levofloxacin lev, by Merck Group (1 mentions)
Anti iκb α antibody, by Santa Cruz Biotechnology (1 mentions)
18 protocols using reagents for cell culture
1
MTT Assay for Cell Viability
2
Recombinant Spider Silk Protein Production
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Unless otherwise stated, all chemicals were of ACS grade, purchased from Sigma-Aldrich Chemie GmbH (Schnelldorf, Germany) and used as supplied. Reagents for cell culture were purchased from Invitrogen (Carlsbad, CA, USA), unless otherwise noted. Human mesenchymal stem cells (HMSCs) were purchased from Lonza Cologne GmbH (Cologne, Germany). High glucose Dulbecco’s Modified Eagle Medium (DMEM) and fetal bovine serum (FBS) were purchased from Biochrom AG (Berlin, Germany). The recombinantly produced silk protein was based on the consensus motif of the repetitive core domain of one of the major ampullate silk fibroins of the garden cross spider (A. diadematus fibroin 4). The recombinant protein is composed of sixteen repeats of the polypeptide module C (amino acid sequence: GSSAAAAAAAASGPGGYGPENQGPSGPGGYGPGGP) and is referred to hereafter as eADF4(C16). Production and purification of eADF4(C16) were carried out as described previously [42 (link)].
3
Comparative Analysis of Egg Sources for Cell Culture
4
Cell Culture Experimental Protocols
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Detailed data for every experiment and additional information regarding materials and methods are presented in Supplement 1 . Cell lines were purchased from ATCC, reagents for cell culture were purchased from Thermo Fisher Scientific, Poland, and all chemical compounds were purchased from Sigma, Poland, unless otherwise indicated.
5
Cell Culture Protocol for Comparative Analysis
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The methods used in this study were previously extensively described in our previous publication8 (link). Therefore, only a brief description is provided in the Material and Methods sections wherever possible. Detailed information is present in “Supplementary materials ”. Cell lines were purchased from ATCC, reagents for cell culture were purchased from Thermo Fisher Scientific, Poland, and all chemical compounds were purchased from Sigma, Poland, unless otherwise indicated.
6
Neuroglioma H4 Cell Culture
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We purchased neuroglioma H4 cells (ATCC® code HTB-148) from LGC Standards (Sesto San Giovanni, Italy). We obtained plasticware from Corning (NY, USA) and reagents for cell culture from Thermo Fisher Scientific (Waltham, MA, USA). If not differently stated, we purchased other reagents from Sigma-Aldrich (St. Louis, MO, USA).
7
Porcine Skin Cell Culture Protocol
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Cell lines were purchased from ATCC, reagents for cell culture were purchased from Thermo Fisher Scientific, Poland, and all chemical compounds were purchased from Sigma, Poland.
Porcine skin was purchased from Stellen Medical LLC, USA.
Porcine skin was purchased from Stellen Medical LLC, USA.
8
Plasmid Transfection and Molecular Assays
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The expression plasmids pcDNA3.1-NFAT2 and its empty vector pcDNA3.1 were purchased from promega (WI, USA). The Lipofectamine 2000 was obtained from Invitrogen (Thermofisher Scientific., USA). Pluronic F-127 was bought from Genecopoeia (MA, USA). Fluo-4/AM, anhydrous dimethyl sulfoxide (DMSO) and cell counting kit-8 (CCK-8) were acquired from Dojindo Molecular Technologies Inc. (Tokyo, Japan). Antibodies against NFAT2 (RRID: AB_2152507), Egr2 (RRID:AB_1139730), FasL (RRID:AB_302235), ERK (RRID:AB_11157324) and β-actin (RRID:AB_764434) were obtained from Abcam (Cambridge, UK). Antibodies against AKT (RRID:AB_329827), p-AKT(Try326) (RRID:AB_1264114), p-ERK(S189) (RRID:AB_490903), c-myc (RRID:AB_2151827) and Cox-2 (RRID:AB_2084968) were bought from Cell Signaling Technology (MA, USA). The HRP-conjugated secondary antibody was acquired from Bioss, Inc. (Beijing, China). Annexin V-FITC apoptosis detection kit was obtained from BD Biosciences (NJ, USA). The Click-iT Plus 5-ethynyl-2′-deoxyuridine (EdU) Flow Cytometry Assay kit was bought from Invitrogen. All chemical reagents used in this study were of analytical grade and all reagents for cell culture were purchased from Gibco BRL Life Technologies (MD, USA).
9
AAV Vector-Based Gene Expression Protocol
10
Anti-inflammatory activity of natural compounds
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ATG, SPA, CLA, CGA, arctiin, deoxycholic acid, glycocholic acid and deoxyglycocholic acid were obtained from Yifang S&T (Tianjin, China). Enzyme-linked immunosorbent assay (ELISA) assay kits of rat TNF-α, IL-6, IL-1β and RANTES, as well as human intracellular proteins (total p38, JNK and ERK), were obtained from Pierce/Endogen Co. (Rockford, Illinois, USA). Human TNF-α was purchased from PeproTech (Rocky Hill, NJ, USA). Dexamethasone (Dex) and levofloxacin (LEV) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Reagents for cell culture were obtained from Gibco BRL Life Technologies (Rockville, MD, USA). The NF-κB luciferase reporter plasmid pGL4.32 and Renilla luciferase reporter vector plasmid pRL-TK were obtained from Promega Co. (Fitchburg, WI, USA). The Lipofectamine 2000 transfection reagent was obtained from Invitrogen (Carlsbad, CA, USA). The anti-IκB-α antibody was obtained from Santa Cruz Biotechnology (San Diego, CA, USA). The P. aeruginosa PAK strain, a clinical isolate from the sputum of a patient suffering from bronchiectasis, was provided by Professor Mingqiang Qiao (College of Life Science, Nankai University, China). All other reagents used in this study were of analytical grade.
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