Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

2.7. Cytotoxicity assay

TB Tetyana P. Buzhdygan
BD Brandon J. DeOre
AB Abigail Baldwin-Leclair
TB Trent A. Bullock
HM Hannah M. McGary
JK Jana A. Khan
RR Roshanak Razmpour
JH Jonathan F. Hale
PG Peter A. Galie
RP Raghava Potula
AA Allison M. Andrews
SR Servio H. Ramirez
ask Ask a question
Favorite

The LIVE/DEAD viability/cytotoxicity assay (Life Technologies, Cat No L3224) was used to evaluate the toxicity of the SARS-CoV-2 spike protein to hBMVECs. Briefly, hBMVECs were seeded on a sterile 96-well plate at 1 × 104 cells per well and grown to confluency. Confluent cells were treated with 10 nM SARS-CoV subunit S1, SARS-CoV-2 subunit S2 or SARS-CoV-2 RBD for 48 h. 200 μL of 1 μM calcein-AM and 5 μM ethidium homodimer-1 were added to each well and incubated for 30 min at room temperature. Data was acquired at excitation and emission wavelengths of 495/515 nm for live cells and 528/617 nm for dead cells and normalized to the total number of cells.

Copyright and License information:  ©2020 The Author(s)
Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A