Dulbecco’s modified Eagle’s medium (DMEM), GlutaMAX, sodium pyruvate and non-essential amino acids were obtained from Life Technologies. Fatty acid-free, ultra-pure BSA and protease inhibitor cocktail were obtained from Roche. BCS, copper chloride, 8-bromo-cAMP, dibutyryl cAMP, IBMX, isoproterenol, and free glycerol reagent were obtained from Sigma-Aldrich. BDH Aristar Ultra concentrated nitric acid was obtained from VWR. NEFA kits were obtained from Wako. H-89 dihydrochloride, cilostamide, and rolipram were obtained from EMD Biosciences. CAY10499 and cyclic AMP assay kits were obtained from Cayman Chemical. Restriction endonucleases, Phusion High-Fidelity DNA polymerase, T4 DNA ligase, and Gibson Assembly Master Mix were obtained from New England Biolabs. TaKaRa ExTaq DNA polymerase was purchased from Clontech. Primers were purchased from Integrated DNA Technologies. All other materials used in biochemical assays were purchased from Sigma unless otherwise noted. Baculoviral particles were generated using the BaculoGold™ transfection kit from BD Biosciences. All DNA constructs were verified by sequencing by Quintara Biosciences, and all plasmids used for transfections were prepared using the EndoFree Maxi Kit (Qiagen).
Baculogold transfection kit
Manufactured by BD
Sourced in United States
The BaculoGold™ transfection kit is a product designed for the transfection of insect cells. It provides the necessary components for the efficient delivery of genetic material into the target cells.
Automatically generated - may contain errors
Lab products found in correlation
Copper chloride, by Merck Group (2 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (2 mentions)
Isoproterenol, by Merck Group (2 mentions)
Dibutyryl camp, by Merck Group (2 mentions)
Endofreemaxi kit, by Qiagen (2 mentions)
Bovine calf serum (bcs), by Merck Group (2 mentions)
8 bromo camp, by Merck Group (2 mentions)
Bdh aristar ultra concentrated nitric acid, by Avantor (2 mentions)
Nefa kit, by Fujifilm (2 mentions)
Cay10499, by Cayman Chemical (2 mentions)
Cyclic amp assay kit, by Cayman Chemical (2 mentions)
Takara ex taq dna polymerase, by Takara Bio (2 mentions)
Gibson assembly master mix, by New England Biolabs (2 mentions)
3 isobutyl 1 methyl xanthine (ibmx), by Merck Group (2 mentions)
Free glycerol reagent, by Merck Group (2 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions)
Phusion high fidelity dna polymerase, by New England Biolabs (2 mentions)
Protease inhibitor cocktail, by Roche (2 mentions)
T4 dna ligase, by New England Biolabs (2 mentions)
Restriction endonuclease, by New England Biolabs (2 mentions)
4 protocols using baculogold transfection kit
1
Analysis of Lipolysis Signaling Pathway
2
Production and Purification of Recombinant rbFcRn
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Recombinant baculoviruses carrying the cDNAs of the soluble rbFcRn α-chain and rbβ2m were generated by using the BaculoGold Transfection Kit (BD Biosciences Pharmingen, Franklin Lakes, NJ, USA) according to the manufacturer’s instructions. Then, Sf9 cells were cultured and infected by recombinant viruses. Recombinant rbFcRn was purified from supernatants of virus infected cells by using Ni-NTA chromatography (His-Select Nickel Affinity Gel, Sigma-Aldrich, St. Louis, MO, USA) according to the manufacturer’s protocol. Expression level and purity of the recombinant protein was confirmed by 15% SDS-PAGE followed by either Coomassie staining or Western blotting. Mouse anti-His tag antibody (AbD Serotec, Kidlington, UK) as the primary (500x) and HRP-conjugated goat anti-mouse IgG (Southern Biotech, Birmingham, AL, USA) as the secondary antibody (10.000x) were used for immunodetection. The yield of the rbFcRn expression was approximately 5 mg/l (5 mg purified protein per 1 liter cell culture supernatant).
3
Baculovirus-mediated OATP Expression in Sf9 Cells
4
Analysis of Lipolysis Signaling Pathway
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Dulbecco’s modified Eagle’s medium (DMEM), GlutaMAX, sodium pyruvate and non-essential amino acids were obtained from Life Technologies. Fatty acid-free, ultra-pure BSA and protease inhibitor cocktail were obtained from Roche. BCS, copper chloride, 8-bromo-cAMP, dibutyryl cAMP, IBMX, isoproterenol, and free glycerol reagent were obtained from Sigma-Aldrich. BDH Aristar Ultra concentrated nitric acid was obtained from VWR. NEFA kits were obtained from Wako. H-89 dihydrochloride, cilostamide, and rolipram were obtained from EMD Biosciences. CAY10499 and cyclic AMP assay kits were obtained from Cayman Chemical. Restriction endonucleases, Phusion High-Fidelity DNA polymerase, T4 DNA ligase, and Gibson Assembly Master Mix were obtained from New England Biolabs. TaKaRa ExTaq DNA polymerase was purchased from Clontech. Primers were purchased from Integrated DNA Technologies. All other materials used in biochemical assays were purchased from Sigma unless otherwise noted. Baculoviral particles were generated using the BaculoGold™ transfection kit from BD Biosciences. All DNA constructs were verified by sequencing by Quintara Biosciences, and all plasmids used for transfections were prepared using the EndoFree Maxi Kit (Qiagen).
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