A replication incompetent pseudotyped HIV (strain SF162) and HIV pseudotyped with GP1,2 of EBOV expressing the firefly luciferase reporter protein (Fluc) were generated as previously described34 (link). In brief, the lentivirus-based pHIV–ZGP–Fluc construct carrying the EBOV glycoprotein GP gene was generated by the co-transfection of 293T cells with pCDNA3.1–EBOV-ZGP-8A and pSG3.Δenv.cmv.Fluc in a 1:2 ratio, using Lipofectamine 3000 (Invitrogen). After incubation for 48 h, the culture supernatant was centrifuged at 210 × g for 5 min, filtered through a 0.45 μM pore-size filter, and concentrated with a 30-kDa ultrafiltration centrifugal tube (Millipore, Boston, MA, USA). All works involving pseudotyped EBOV were performed in a BSL-2 facility at the National Institutes of Food and Drug Control, Beijing, China.
30 kda ultrafiltration centrifugal tube
Manufactured by Merck Group
Sourced in United States
The 30-kDa ultrafiltration centrifugal tube is a laboratory equipment designed for the separation and concentration of macromolecules, such as proteins, based on their molecular weight. The tube utilizes a semi-permeable membrane with a molecular weight cutoff of 30 kilodaltons, allowing for the selective filtration of molecules smaller than this threshold while retaining larger molecules during a centrifugation process.
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Lab products found in correlation
2 protocols using 30 kda ultrafiltration centrifugal tube
1
Pseudotyped HIV and EBOV Lentivirus Production
2
Generation of Ebola and Marburg Pseudotyped Viruses
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HIV pseudotyped viruses with GP1,2 of EBOV, GP of MARV expressing the firefly luciferase reporter protein (Fluc) were generated as previously described21 (link), 24 (link). In brief, the lentivirus-based pHIV-EBOVGP-Fluc construct carrying the EBOV GP gene, and pHIV-MARVGP-Fluc construct carrying the MARV GP gene were generated by the co-transfection of 293T cells with pCDNA3.1-EBOV-ZGP-8A and pCDNA3.1-MARVGP respectively, together and HIV-1 containing firefly luciferase reporter gene (pSG3.cmv.Fluc) in a 1:2 ratio using Lipofectamine 3000 (Invitrogen). After incubation for 48 h, the culture supernatant was centrifuged at 210 × g for 5 min, filtered through a 0.45 μmol/L pore-size filter, and concentrated with a 30-kDa ultrafiltration centrifugal tube (Millipore, Boston, MA, USA). All works involving pseudotyped virus were performed in a BSL-2 facility at the National Institutes of Food and Drug Control, Beijing, China.
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