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10k amicon tubes

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The 10K Amicon tubes are centrifugal filter devices designed for the concentration and purification of macromolecules, including proteins, peptides, and other biological samples. These tubes feature a 10,000 molecular weight cut-off (MWCO) membrane, which allows the passage of smaller molecules while retaining the desired larger molecules.

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Lab products found in correlation

Econo gradient pump, by Bio-Rad (4 mentions) Expi293f, by Thermo Fisher Scientific (1 mentions)

4 protocols using 10k amicon tubes

1

Fab Generation from IgG

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To generate the Fab from the IgG, a stop codon was inserted in the heavy chain constant region at “KSCDK”. The truncated heavy chains were co-transfected with the corresponding light chains in 293Expi cells to produce the Fabs. The supernatants were harvested 4 days post transfection. Fabs were purified with CaptureSelect™ CH1-XL MiniChrom Columns (#5943462005). Supernatants were loaded onto columns using an Econo Gradient Pump (Bio-Rad #7319001). Following a wash with 1x PBS, Fabs were eluted with 25 mL of 50 mM acetate (pH 4.0) and neutralized with 2 M Tris Base. The eluate was buffer exchanged with 1x PBS in 10K Amicon tubes (Millipore, UFC901008) and filtered with a 0.22 μm spin filter.
He W.T., Musharrafieh R., Song G., Dueker K., Tse L.V., Martinez D.R., Schäfer A., Callaghan S., Yong P., Beutler N., Torres J.L., Volk R.M., Zhou P., Yuan M., Liu H., Anzanello F., Capozzola T., Parren M., Garcia E., Rawlings S.A., Smith D.M., Wilson I.A., Safonova Y., Ward A.B., Rogers T.F., Baric R.S., Gralinski L.E., Burton D.R, & Andrabi R. (2022). Targeted isolation of panels of diverse human protective broadly neutralizing antibodies against SARS-like viruses. bioRxiv.
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2

Fab Production from IgG Truncation

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To generate the Fab from the IgG, a stop codon was inserted in the heavy chain constant region at “KSCDK”. The truncated heavy chains were co-transfected with the corresponding light chains in Expi293F (Life Technologies) cells to produce the Fabs. The supernatants were harvested 4 days post transfection. Fabs were purified with CaptureSelect™ CH1-XL MiniChrom Columns (#5943462005). Supernatants were loaded onto columns using an Econo Gradient Pump (Bio-Rad #7319001). Following a wash with 1x PBS, Fabs were eluted with 25 mL of 50 mM acetate (pH 4.0) and neutralized with 2 M Tris Base. The eluate was buffer exchanged with 1x PBS in 10K Amicon tubes (Millipore, UFC901008) and filtered with a 0.22 μm spin filter.
He W.T., Musharrafieh R., Song G., Dueker K., Tse L.V., Martinez D.R., Schäfer A., Callaghan S., Yong P., Beutler N., Torres J.L., Volk R.M., Zhou P., Yuan M., Liu H., Anzanello F., Capozzola T., Parren M., Garcia E., Rawlings S.A., Smith D.M., Wilson I.A., Safonova Y., Ward A.B., Rogers T.F., Baric R.S., Gralinski L.E., Burton D.R, & Andrabi R. (2022). Targeted isolation of panels of diverse human protective broadly neutralizing antibodies against SARS-like viruses. Nature immunology, 23(6), 960-970.
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3

Fab Production from Truncated IgG

Check if the same lab product or an alternative is used in the 5 most similar protocols
To generate the Fab of an IgG, a stop codon was inserted in the heavy chain constant region at “KSCDK*.” The truncated heavy chains were cotransfected with the corresponding light chains in Expi293 cells to produce the Fab. The supernatants were harvested 4 days after transfection. Fabs were purified with CaptureSelect CH1-XL MiniChrom Columns (#5943462005). Supernatants were loaded onto columns using an Econo Gradient Pump (Bio-Rad, #7319001). After a wash with 1× PBS, Fabs were eluted with 25 ml of 50 mM acetate (pH 4.0) and neutralized with 2 M tris base. The eluate was buffer-exchanged with 1× PBS in 10K Amicon tubes (Millipore, UFC901008) and filtered with a 0.22-μm spin filter.
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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4

Fab Production from Truncated IgG

Check if the same lab product or an alternative is used in the 5 most similar protocols
To generate the Fab of an IgG, a stop codon was inserted in the heavy chain constant region at “KSCDK*.” The truncated heavy chains were cotransfected with the corresponding light chains in Expi293 cells to produce the Fab. The supernatants were harvested 4 days after transfection. Fabs were purified with CaptureSelect CH1-XL MiniChrom Columns (#5943462005). Supernatants were loaded onto columns using an Econo Gradient Pump (Bio-Rad, #7319001). After a wash with 1× PBS, Fabs were eluted with 25 ml of 50 mM acetate (pH 4.0) and neutralized with 2 M tris base. The eluate was buffer-exchanged with 1× PBS in 10K Amicon tubes (Millipore, UFC901008) and filtered with a 0.22-μm spin filter.
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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