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Steriflip sterile disposable vacuum filter units

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The Steriflip Sterile Disposable Vacuum Filter Units are a type of laboratory equipment designed for the filtration of liquids. They provide a sterile environment for the filtration process, ensuring the integrity of the sample.

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Lab products found in correlation

Hek293f cell, by Thermo Fisher Scientific (2 mentions) Expi293 cell, by Thermo Fisher Scientific (2 mentions) Fectopro reagent, by Polyplus Transfection (2 mentions) Transfectagro, by Corning (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Hek293t cell, by American Type Culture Collection (1 mentions)

Close spelling variants of this product

Steriflip filter unit Steriflip unit Steriflip filter Filter units Disposable filter Vacuum filters Sterile filters Steriflip

3 protocols using steriflip sterile disposable vacuum filter units

1

Transient Expression and Purification of Antibodies and Viral Proteins

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To express Abs, the corresponding heavy chain and light chain plasmids were transiently transfected into the Expi293 cell (Life Technologies) at 3 × 106 cells/ml with FectoPRO PolyPlus transfection reagent (Polyplus, catalog no. 116–040). To boost the protein yield, we fed cells with 300 mM sodium valproic acid solution and glucose solution 1 day after transfection. To purify mAbs, the supernatants were harvested 4 days after transfection. To express the soluble S ectodomain proteins from SARS-CoV-1, SARS-CoV-2, and their truncations, plasmids were transfected into human embryonic kidney (HEK) 293F cells (Life Technologies) at 1 million cells/ml. For 1 liter of HEK293F transfection, we combined 350 μg of plasmids with 16 ml of transfectagro (Corning) and filtered with 0.22-μm Steriflip Sterile Disposable Vacuum Filter Units (Millipore Sigma). The 1.8 ml of 40K polyethylenimine (PEI) (1mg/ml) was mixed into 16 ml of transfectagro in another conical tube. The filtered plasmid solution was gently combined with the mixed PEI solution by inverting the tube several times. After resting at room temperature for 30 min, the mixture was poured into 1 liter of HEK293F cells. The supernatant was centrifuged and filtered with 0.22-μm membrane to a glass bottle 5 days after transfection.
He W.T., Yuan M., Callaghan S., Musharrafieh R., Song G., Silva M., Beutler N., Lee W.H., Yong P., Torres J.L., Melo M., Zhou P., Zhao F., Zhu X., Peng L., Huang D., Anzanello F., Ricketts J., Parren M., Garcia E., Ferguson M., Rinaldi W., Rawlings S.A., Nemazee D., Smith D.M., Briney B., Safonova Y., Rogers T.F., Dan J.M., Zhang Z., Weiskopf D., Sette A., Crotty S., Irvine D.J., Ward A.B., Wilson I.A., Burton D.R, & Andrabi R. (2022). Broadly neutralizing antibodies to SARS-related viruses can be readily induced in rhesus macaques. Science translational medicine, 14(657), eabl9605.
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2

Pseudovirus Generation for Virus Spike Proteins

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The spike proteins of each tested virus were cloned into a plasmid expression vector with each protein’s endoplasmic reticulum retrieval signal removed. These plasmids were co-transfected with MLV (murine leukemia virus)-CMV (cytomegalovirus) luciferase and MLV Gag/Pol plasmids into HEK-293T cells (ATCC CRL-3216) using Lipofectamine 2000 (Thermo Fisher 11668019) transfection reagent following the manufacturer’s recommended protocol. 16 hours after transfection, cell media was replaced with fresh warm cell media (DMEM (Corning 10–017-CV) with 10% FBS (Omega Scientific NC0471611), 1% L-glutamine (Corning 25–005-CI), and 1% penicillin-streptomycin (Corning 30–002-CI)). 48 hours after transfection, supernatant was collected and filtered using 0.22 μm Steriflip™ Sterile Disposable Vacuum Filter Units (MilliporeSigma™ SCGP00525) and the resulting pseudoviruses were stored at −80 °C for later use.
Song G., Yuan M., Liu H., Capozzola T., Lin R.N., Torres J.L., He W.T., Musharrafieh R., Dueker K., Zhou P., Callaghan S., Mishra N., Yong P., Anzanello F., Avillion G., Vo A.L., Li X., Makhdoomi M., Feng Z., Zhu X., Peng L., Nemazee D., Safonova Y., Briney B., Ward A.B., Burton D.R., Wilson I.A, & Andrabi R. (2023). Broadly neutralizing antibodies targeting a conserved silent face of spike RBD resist extreme SARS-CoV-2 antigenic drift. bioRxiv.
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3

Transient Expression and Purification of Antibodies and Viral Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
To express Abs, the corresponding heavy chain and light chain plasmids were transiently transfected into the Expi293 cell (Life Technologies) at 3 × 106 cells/ml with FectoPRO PolyPlus transfection reagent (Polyplus, catalog no. 116–040). To boost the protein yield, we fed cells with 300 mM sodium valproic acid solution and glucose solution 1 day after transfection. To purify mAbs, the supernatants were harvested 4 days after transfection. To express the soluble S ectodomain proteins from SARS-CoV-1, SARS-CoV-2, and their truncations, plasmids were transfected into human embryonic kidney (HEK) 293F cells (Life Technologies) at 1 million cells/ml. For 1 liter of HEK293F transfection, we combined 350 μg of plasmids with 16 ml of transfectagro (Corning) and filtered with 0.22-μm Steriflip Sterile Disposable Vacuum Filter Units (Millipore Sigma). The 1.8 ml of 40K polyethylenimine (PEI) (1mg/ml) was mixed into 16 ml of transfectagro in another conical tube. The filtered plasmid solution was gently combined with the mixed PEI solution by inverting the tube several times. After resting at room temperature for 30 min, the mixture was poured into 1 liter of HEK293F cells. The supernatant was centrifuged and filtered with 0.22-μm membrane to a glass bottle 5 days after transfection.
He W.T., Yuan M., Callaghan S., Musharrafieh R., Song G., Silva M., Beutler N., Lee W.H., Yong P., Torres J.L., Melo M., Zhou P., Zhao F., Zhu X., Peng L., Huang D., Anzanello F., Ricketts J., Parren M., Garcia E., Ferguson M., Rinaldi W., Rawlings S.A., Nemazee D., Smith D.M., Briney B., Safonova Y., Rogers T.F., Dan J.M., Zhang Z., Weiskopf D., Sette A., Crotty S., Irvine D.J., Ward A.B., Wilson I.A., Burton D.R, & Andrabi R. (2022). Broadly neutralizing antibodies to SARS-related viruses can be readily induced in rhesus macaques. Science translational medicine, 14(657), eabl9605.
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