Dulbecco’s modified minimal essential medium (DMEM), fetal bovine serum (FBS), phosphate-buffered saline (PBS), Trypsin-EDTA solution, 100× penicillin–streptomycin–l -glutamine solution, anti-RFP antibody, Alexa 647-conjugated secondary antibody, SlowFade Gold antifade reagent, and a LIVE/DEAD fixable yellow dead cell stain kit were purchased from Thermo Fisher (Waltham, MA USA). Puromycin solution and G418 solution were obtained from InvivoGen (San Diego, CA, USA). Hygromycin B solution was purchased from Enzo Life Science (Farmingdale, NY, USA). Anti-ORF57, anti-K8, and anti-K8.1 antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Anti-LANA and anti-lamin A/C antibodies were purchased from Millipore-Sigma (St. Louis, MO, USA). Anti-nucleolin antibody was purchased from Abcam (Cambridge, UK). Anti-K-Rta antibody was described previously (68 (link)). Monoclonal anti-ORF52 antibody was a generous gift from Fanxiu Zhu (Florida State University). All other chemicals were purchased from Millipore-Sigma (St. Louis, MO, USA) unless otherwise stated.
Puromycin solution
Manufactured by InvivoGen
Sourced in France
Puromycin solution is a laboratory reagent used for the selection and maintenance of cells that express a puromycin resistance gene. It inhibits protein synthesis in eukaryotic cells, allowing for the selection of successfully transfected or transduced cells.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Restriction endonuclease dpni, by Thermo Fisher Scientific (2 mentions)
Turbofect cationic polymerbased transfection reagent, by Thermo Fisher Scientific (2 mentions)
Genejet dna spin column based purification kit, by Thermo Fisher Scientific (2 mentions)
Endotoxin binding reagent, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (2 mentions)
Alexa 647 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Slowfade gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Trypsin edta solution, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Live dead fixable yellow dead cell stain kit, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified minimal essential medium, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin l glutamine solution, by Thermo Fisher Scientific (1 mentions)
Anti rfp antibody, by Thermo Fisher Scientific (1 mentions)
Hygromycin b solution, by Enzo Life Sciences (1 mentions)
Anti orf57, by Santa Cruz Biotechnology (1 mentions)
Anti k8, by Santa Cruz Biotechnology (1 mentions)
Anti nucleolin antibody, by Abcam (1 mentions)
Ampicillin sodium salt, by Thermo Fisher Scientific (1 mentions)
Milli q purification system, by Merck Group (1 mentions)
3 protocols using puromycin solution
1
Immunofluorescence Staining Protocol
2
Production and Purification of SMALPs
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Commercial styrene-maleic acid (SMAc) co-polymers X25 (Xiran SL 25010 P20, lot number from EtOH before use. THF was purified and dried according to the reported procedures and stored under nitrogen [22] (link). MAn was used as received.
Buffers for the preparation of SMALPs were prepared in ultrapure water treated with a Milli-Q purification system (Merck, USA). Potassium phosphate monobasic (KH2PO4), potassium phosphate dibasic (K2HPO4), sodium chloride (NaCl), tris(hydroxymethyl)aminomethane (Tris), hydrochloric acid (HCl), glycerol, sucrose, calcium chloride (CaCl2) dihydrate, protease inhibitor cocktail tablets (SigmaFAST), ethylenediaminetetraacetic acid disodium salt (Na2EDTA), nickel(II) chloride (NiCl2) hexahydrate, imidazole and polyethylene glycol sorbitan monolaurate (Tween-20) were purchased from Sigma-Aldrich (Merck, USA).
The pX459 plasmid was purchased from Addgene (USA). Dulbecco's Modified Eagle Medium (DMEM), fetal bovine serum (FBS), penicillin-streptomycin solution (5000 U/mL penicillin, 5000 µg/mL streptomycin), ampicillin sodium salt, the restriction endonuclease DpnI, the Turbofect cationic polymerbased transfection reagent, the GeneJet DNA spin column-based purification kit and the endotoxin binding reagent were purchased from ThermoFisher Scientific (USA). Puromycin solution (10 mg/mL) was purchased from Invivogen (France).
Buffers for the preparation of SMALPs were prepared in ultrapure water treated with a Milli-Q purification system (Merck, USA). Potassium phosphate monobasic (KH2PO4), potassium phosphate dibasic (K2HPO4), sodium chloride (NaCl), tris(hydroxymethyl)aminomethane (Tris), hydrochloric acid (HCl), glycerol, sucrose, calcium chloride (CaCl2) dihydrate, protease inhibitor cocktail tablets (SigmaFAST), ethylenediaminetetraacetic acid disodium salt (Na2EDTA), nickel(II) chloride (NiCl2) hexahydrate, imidazole and polyethylene glycol sorbitan monolaurate (Tween-20) were purchased from Sigma-Aldrich (Merck, USA).
The pX459 plasmid was purchased from Addgene (USA). Dulbecco's Modified Eagle Medium (DMEM), fetal bovine serum (FBS), penicillin-streptomycin solution (5000 U/mL penicillin, 5000 µg/mL streptomycin), ampicillin sodium salt, the restriction endonuclease DpnI, the Turbofect cationic polymerbased transfection reagent, the GeneJet DNA spin column-based purification kit and the endotoxin binding reagent were purchased from ThermoFisher Scientific (USA). Puromycin solution (10 mg/mL) was purchased from Invivogen (France).
3
CRISPR-Cas9 Plasmid Transfection
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Commercial styrene-maleic acid (SMAc) co-polymers X25 (Xiran SL 25010 P20, lot number The PX459 plasmid (pSpCas9(BB)-2A-Puro, V2.0) was a gift from Feng Zhang (Addgene plasmid #62988) [30] (link). Dulbecco's Modified Eagle Medium (DMEM), phosphate buffered saline (PBS), fetal bovine serum (FBS), penicillin-streptomycin solution (5000 U/mL penicillin, 5000 µg/mL streptomycin), ampicillin sodium salt, the restriction endonuclease DpnI, the Turbofect cationic polymer-based transfection reagent, the GeneJet DNA spin column-based purification kit and the endotoxin binding reagent were purchased from Thermo Fisher Scientific (USA). Puromycin solution (10 mg/mL) was purchased from Invivogen (France).
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