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Econo pak chromatography column

Manufactured by Bio-Rad

The Econo-Pak Chromatography column is a pre-packed, disposable chromatography column designed for small-scale purification and separation of biomolecules. The column is made of durable polypropylene and features a built-in frit to support the chromatography matrix. The column dimensions and matrix can be selected based on the specific application requirements.

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4 protocols using econo pak chromatography column

1

Purification of ECD-Fc Proteins from Expi293F Cells

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ECD-Fc proteins were produced in Expi293F cells using transfection conditions described above. Following harvesting of cell conditioned media, 1 M Tris, pH 8.0 was added to a final concentration of 20 mM. Ni-NTA Agarose (QIAGEN) was added to ∼5% conditioned media volume. 1 M sterile PBS, pH 7.2 (GIBCO) was added to ∼3X conditioned media volume. The mixture was stirred overnight at 4°C. Ni-NTA Agarose beads were collected in a Buchner funnel and washed with ∼300 mL protein wash buffer (30 mM HEPES, pH 7.2, 150 mM NaCl, 20 mM imidazole). Beads were transferred to an Econo-Pak Chromatography column (Bio-Rad) and protein was eluted in 15 mL of elution buffer (30 mM HEPES, pH 7.2, 150 mM NaCl, 200 mM imidazole). Proteins were concentrated using Amicon Ultracel 10K filters (Millipore) to a volume of ∼0.5 mL and absorbance at 280 nm was measured using a Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific) to determine protein concentration.
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2

Recombinant Ly6a-Fc Protein Production

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Ly6a-Fc was produced in Expi293F suspension cells grown in Expi293 Expression Medium (Thermo Fisher Scientific) in a 37 °C, 5% CO2 incubator with 130 rpm shaking. Transfection was performed with Expifectamine according to the manufacturer’s instructions (Thermo Fisher Scientific). Following harvesting of cell-conditioned media, 1 M Tris, pH 8.0, was added to a final concentration of 20 mM. Ni-NTA Agarose (QIAGEN) was added to ∼5% conditioned media volume. 1 M sterile PBS, pH 7.2 (GIBCO), was added to ∼3X conditioned media volume. The mixture was stirred overnight at 4 °C. Ni-NTA Agarose beads were collected in a Buchner funnel and washed with ∼300 mL protein wash buffer (30 mM HEPES, pH 7.2, 150 mM NaCl, 20 mM imidazole). Beads were transferred to an Econo-Pak Chromatography column (Bio-Rad) and protein was eluted in 15 mL of elution buffer (30 mM HEPES, pH 7.2, 150 mM NaCl, and 200 mM imidazole). Proteins were concentrated using Amicon Ultracel 10 K filters (Millipore) and absorbance at 280 nm was measured using a Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific) to determine protein concentration.
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3

Protein Purification from Expi293F Cells

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Proteins were produced in Expi293F cells using transfection conditions following the manufacturer’s protocol. After harvesting of cell media, 1 M Tris, pH 8.0 was added to a final concentration of 20 mM. Ni-NTA Agarose (Qiagen) was added to ~5% media volume. ×1 sterile PBS, pH 7.2 (Gibco) was added to ~×3 media volume. The mixture was stirred overnight at 4°C. Ni-NTA Agarose beads were collected in a Buchner funnel and washed with ~300 ml protein wash buffer (20 mM HEPES, pH 7.2, 150 mM NaCl, 20 mM imidazole). Beads were transferred to an Econo-Pak Chromatography column (Bio-Rad) and protein was eluted in 15 ml of elution buffer (20 mM HEPES, pH 7.2, 150 mM NaCl, 200 mM imidazole). Proteins were concentrated using Amicon Ultracel filters (Millipore) and absorbance at 280 nm was measured using a Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific) to determine protein concentration. A summary of the expression yields can be found in Figure 2—source data 2.
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4

Recombinant Ly6a-Fc Protein Production

Check if the same lab product or an alternative is used in the 5 most similar protocols
Ly6a-Fc was produced in Expi293F suspension cells grown in Expi293 Expression Medium (Thermo Fisher Scientific) in a 37 °C, 5% CO2 incubator with 130 rpm shaking. Transfection was performed with Expifectamine according to manufacturer’s instructions (Thermo Fisher Scientific). Following harvesting of cell conditioned media, 1 M Tris, pH 8.0 was added to a final concentration of 20 mM. Ni-NTA Agarose (QIAGEN) was added to ~5% conditioned media volume. 1 M sterile PBS, pH 7.2 (GIBCO) was added to ~3X conditioned media volume. The mixture was stirred overnight at 4 °C. Ni-NTA Agarose beads were collected in a Buchner funnel and washed with ~300 mL protein wash buffer (30 mM HEPES, pH 7.2, 150 mM NaCl, 20 mM imidazole). Beads were transferred to an Econo-Pak Chromatography column (Bio-Rad) and protein was eluted in 15 mL of elution buffer (30 mM HEPES, pH 7.2, 150 mM NaCl, 200 mM imidazole). Proteins were concentrated using Amicon Ultracel 10K filters (Millipore) and absorbance at 280 nm was measured using a Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific) to determine protein concentration.
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