Recombinant PEDV S1 subunit was prepared as previously described by us [38 (link)]. Briefly, recombinant PEDV S1 subunit was expressed by the Drosophila S2 cells in Sf-900 II serum-free medium (Invitrogen, Carlsbad, USA), and purified by HisTrap excel prepacked column (GE Healthcare, Fairfield, USA) and HiTrap Q HP prepacked column (GE Healthcare). The purified S1 subunit was applied to 15% SDS-PAGE for analysis.
Hitrap q hp prepacked column
Manufactured by GE Healthcare
Sourced in United States
The HiTrap Q HP prepacked column is a lab equipment product designed for anion exchange chromatography. It is a pre-packed column that allows for efficient and convenient purification of biomolecules. The column is made of a strong anion exchanger medium, which enables the capture and separation of negatively charged molecules.
Automatically generated - may contain errors
Lab products found in correlation
Histrap excel prepacked column, by GE Healthcare (2 mentions)
Hitrap, by Cytiva (2 mentions)
Sf900 2, by Thermo Fisher Scientific (1 mentions)
Superdex 200 increase 10 300 gl, by GE Healthcare (1 mentions)
Hiload 16 600 superdex 200 pg, by GE Healthcare (1 mentions)
Human plasminogen, by Merck Group (1 mentions)
Pepstatin, by Merck Group (1 mentions)
Bca protein assay kit, by Solarbio (1 mentions)
3 protocols using hitrap q hp prepacked column
1
Recombinant PEDV S1 Subunit Production
2
Purification and Deglycosylation of Viral Proteins
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The supernatant containing pAPN ectodomain, PEDV S1 or S1t protein was adjusted to pH 8.0 by 1.5 M Tris-HCl pH 8.8, followed by centrifugation at 10000 rpm at 4 °C for 30 min. After filtration through 0.22 μm filter membranes, the supernatant was applied to HisTrap excel prepacked column (GE Healthcare, Fairfield, USA) or/and HiTrap Q HP prepacked column (GE Healthcare) for purification. Then, each target protein was subjected to size-exclusion column Hiload 16/600 Superdex 200 pg (GE Healthcare) and Superdex 200 increase 10/300 GL (GE Healthcare) for further purification with 20 mM Tris-HCl pH 7.6 and 150 mM NaCl as the elution buffer. The purified proteins were treated with PNGase F according to the manufacturer's instructions. The PNGase F-treated and -untreated proteins were applied to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing (with β-mercaptoethanol) or non-reducing condition (without β-mercaptoethanol).
3
Purification and Characterization of Lumbrokinase
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Living earthworms (P. vulgaris) were provided by Shanghai Dilong Breeding Base and identified by Professor Du Shouying at Beijing University of Chinese Medicine. The crude extract of P. vulgaris was obtained by water extraction and alcohol precipitation. Lumbrokinase, fibrinogen, and thrombin were purchased from China National Institutes for Food and Drug Control. BCA Protein Assay Kit was obtained from Solarbio life sciences (China). Human plasminogen, phenylmethane sulfonyl fluoride (PMSF), soybean trypsin inhibitor (SBTI), N--tosyl-l-phenylalanine (TPCK), and pepstatin were purchased from Sigma (United States). Gel filtration low molecular weight marker protein was obtained from Thermo Fisher Scientific (United States). Superdex 75 Increase 10/300 GL pre-packed columns and HiTrap Q HP pre-packed column were purchased from GE life sciences (United States). Other reagents used were of analytical grade.
Sprague–Dawley (SD) rats were purchased from Hunan SLAC Laboratory Animal Co., Ltd. (Changsha, China). All experimental protocols were approved by the Animal Experimentation Committee of the Beijing University of Chinese Medicine (Beijing, China).
Sprague–Dawley (SD) rats were purchased from Hunan SLAC Laboratory Animal Co., Ltd. (Changsha, China). All experimental protocols were approved by the Animal Experimentation Committee of the Beijing University of Chinese Medicine (Beijing, China).
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