293T cells transfected with CoV spike pseudotypes (producer cells) were lysed in 1% sodium dodecyl sulfate, 150mM NaCl, 50 mM Tris-HCl and 5 mM EDTA and centrifuged at 14,000g for 20 minutes. Pseudotyped particles were concentrated from producer cell supernatants that were overlaid on a 10% OptiPrep cushion in PBS (Sigma–Aldrich) and centrifuged at 20,000g for 2h at 4 °C. Lysates and concentrated particles were analyzed for FLAG (Sigma–Aldrich; A8592; 1:10,000), GAPDH (Sigma–Aldrich; G8795; 1:10,000) and/or VSV-M (Kerafast; 23H12; 1:5,000) expression on 10% Bis-Tris PAGE gel (Thermo Fisher Scientific).
G8795
Manufactured by Kerafast
G8795 is a laboratory instrument used for performing electrophoretic separations. It is designed to separate and analyze various biomolecules, such as proteins, nucleic acids, or small molecules, based on their size and charge characteristics.
Automatically generated - may contain errors
3 protocols using g8795
1
Purification and Analysis of SARS-CoV-2 Spike Pseudotypes
2
Pseudotyped Particle Production and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Producer cells (spike-transfected 293T) were lysed in 1% sodium dodecyl sulfate, 150 mM NaCl, 50 mM Tris-HCl and 5 mM EDTA and clarified by centrifugation at 14,000g for 20 min.
Pseudotyped particles were concentrated from producer cell supernatants that were overlaid on a 10% OptiPrep cushion in PBS (Sigma–Aldrich) and centrifuged at 20,000g for 2 h at 4 °C. Lysates and concentrated particles were analysed for FLAG (Sigma–Aldrich; A8592; 1:10,000), GAPDH (Sigma–Aldrich; G8795; 1:10,000) and/or VSV-M (Kerafast; 23H12; 1:5,000) expression on 10% Bis-Tris PAGE gel (Thermo Fisher Scientific).
Pseudotyped particles were concentrated from producer cell supernatants that were overlaid on a 10% OptiPrep cushion in PBS (Sigma–Aldrich) and centrifuged at 20,000g for 2 h at 4 °C. Lysates and concentrated particles were analysed for FLAG (Sigma–Aldrich; A8592; 1:10,000), GAPDH (Sigma–Aldrich; G8795; 1:10,000) and/or VSV-M (Kerafast; 23H12; 1:5,000) expression on 10% Bis-Tris PAGE gel (Thermo Fisher Scientific).
3
Purification and Analysis of Pseudotyped Viral Particles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
293T cells transfected with CoV spike pseudotypes (producer cells) were lysed in 1% sodium dodecyl sulfate, 150 mM NaCl, 50 mM Tris-HCl, and 5 mM EDTA and centrifuged at 14,000g for 20 minutes. Pseudotyped particles were concentrated from producer cell supernatants that were overlaid on a 10% OptiPrep cushion in PBS (Sigma–Aldrich) and centrifuged at 20,000g for 2 h at 4°C. Lysates and concentrated particles were analyzed for FLAG (Sigma–Aldrich; A8592; 1:10,000), GAPDH (Sigma–Aldrich; G8795; 1:10,000), and/or VSV-M (Kerafast; 23H12; 1:5,000) expression on 10% Bis-Tris PAGE gel (Thermo Fisher Scientific).
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