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Vivaspin 30 kda mwco filters

Manufactured by Sartorius
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The Vivaspin 30 kDa MWCO filters are a type of centrifugal concentrator used for the separation and concentration of macromolecules, such as proteins, from solutions. These filters have a molecular weight cut-off (MWCO) of 30 kDa, which means they are designed to retain molecules larger than 30 kDa while allowing smaller molecules to pass through.

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Lab products found in correlation

Endo hf, by New England Biolabs (3 mentions) His tag antibody, by Thermo Fisher Scientific (3 mentions) Sonicated, by Qsonica (3 mentions) Glycobuffer 3, by New England Biolabs (3 mentions) Nebexpress ni spin columns, by New England Biolabs (3 mentions)

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Vivaspin (140 citations) Vivaspin filters (26 citations)

3 protocols using vivaspin 30 kda mwco filters

1

Purification and Glycosylation Analysis of Recombinant Proteins

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Example 4

The spent culture media (P. pastoris constructs) were buffer-exchanged in 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and concentrated ten-fold using Vivaspin 30 kDa MWCO filters (Sartorius). To assess the extent of glycosylation, the concentrated spent culture media were subject to Endo Hf (New England Biolabs) digestions under native conditions in the presence of 1× GlycoBuffer 3 (50 mM sodium acetate, pH 6.0).

FCE proteins (wild-type and N-glycan variants) were purified from the concentrated spent cultures using NEBExpress Ni Spin Columns (New England Biolabs). The columns were washed twice with 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and 5 mM imidazole then once with 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and 10 mM imidazole. The purified protein was eluted with 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and 500 mM imidazole.

To prepare cell lysates (K. lactis constructs), cells were resuspended in 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and sonicated (Qsonica). The soluble cell lysate was pre-cleared by centrifugation at 16000×g for 15 minutes at 4° C. The cell lysates and spent culture media were analyzed by SDS-PAGE on 4-20% polyacrylamide gel, followed by western blotting with a His-tag antibody (Thermo Fisher Scientific).

US11098295B1. FCE mRNA capping enzyme compositions, methods and kits (2021-08-24). New England Biolabs, Inc. [US]. Inventors: Mehul Ganatra [US], Siu-Hong Chan [US], Christopher H. Taron [US], G. B. Robb [US].
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2

Purification and Analysis of P. pastoris and K. lactis Proteins

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Example 4

The spent culture media (P. pastoris constructs) were buffer-exchanged in 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and concentrated ten-fold using Vivaspin 30 kDa MWCO filters (Sartorius). To assess the extent of glycosylation, the concentrated spent culture media were subject to Endo Hf (New England Biolabs) digestions under native conditions in the presence of 1× GlycoBuffer 3 (50 mM sodium acetate, pH 6.0).

FCE proteins (wild-type and N-glycan variants) were purified from the concentrated spent cultures using NEBExpress Ni Spin Columns (New England Biolabs). The columns were washed twice with 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and 5 mM imidazole then once with 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and 10 mM imidazole. The purified protein was eluted with 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and 500 mM imidazole.

To prepare cell lysates (K. lactis constructs), cells were resuspended in 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and sonicated (Qsonica). The soluble cell lysate was pre-cleared by centrifugation at 16000×g for 15 minutes at 4° C. The cell lysates and spent culture media were analyzed by SDS-PAGE on 4-20% polyacrylamide gel, followed by western blotting with a His-tag antibody (Thermo Fisher Scientific).

US11028379B1. FCE mRNA capping enzyme compositions, methods and kits (2021-06-08). New England Biolabs, Inc. [US]. Inventors: Mehul Ganatra [US], Siu-hong Chan [US], Christopher H. Taron [US], G. B. Robb [US].
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3

Purification and Analysis of P. pastoris and K. lactis Constructs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 4

The spent culture media (P. pastoris constructs) were buffer-exchanged in 50 mM Tris-HC1, pH 7.5 buffer containing 300 mM NaCl and concentrated ten-fold using Vivaspin 30 kDa MWCO filters (Sartorius). To assess the extent of glycosylation, the concentrated spent culture media were subject to Endo Hf (New England Biolabs) digestions under native conditions in the presence of 1× GlycoBuffer 3 (50 mM sodium acetate, pH 6.0).

FCE proteins (wild-type and N-glycan variants) were purified from the concentrated spent cultures using NEBExpress Ni Spin Columns (New England Biolabs). The columns were washed twice with 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and 5 mM imidazole then once with 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and 10 mM imidazole. The purified protein was eluted with 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and 500 mM imidazole.

To prepare cell lysates (K. lactis constructs), cells were resuspended in 50 mM Tris-HCl, pH 7.5 buffer containing 300 mM NaCl and sonicated (Qsonica). The soluble cell lysate was pre-cleared by centrifugation at 16000×g for 15 minutes at 4° C. The cell lysates and spent culture media were analyzed by SDS-PAGE on 4-20% polyacrylamide gel, followed by western blotting with a His-tag antibody (Thermo Fisher Scientific).

US11725196B2. FCE mRNA capping enzyme compositions, methods and kits (2023-08-15). New England Biolabs, Inc. [US]. Inventors: Mehul Ganatra [US], Siu-hong Chan [US], Christopher H. Taron [US], G. B. Robb [US].
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