Blocking activity of purified rabbit anti-RBD antibodies on the binding of RBD proteins with a His tag to 293T/ACE2 cells was detected by flow cytometry assay. Briefly, SARS-CoV-2 RBD protein (2 μg/ml) or SARS-CoV RBD protein (10 μg/ml) was added to 4 × 105 cells and incubated for 30 min at room temperature. After washing two times with PBS, cells were incubated with a 1:500 dilution of Alexa Fluor 488–labeled rabbit anti–His tag antibody (Cell Signaling Technology, Danvers, MA) for 30 min at room temperature. After two washes, cells were resuspended in PBS and analyzed by FACSCantoII instrument (Becton Dickinson, Mountain View, CA).
Alexa fluor 488 labeled rabbit anti his tag antibody
Manufactured by Cell Signaling Technology
Sourced in United States
Alexa Fluor 488–labeled rabbit anti–His tag antibody is a fluorescently-labeled antibody that recognizes the polyhistidine (His) tag. It is used to detect and visualize proteins that have been tagged with a His sequence.
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2 protocols using alexa fluor 488 labeled rabbit anti his tag antibody
1
SARS-CoV-2 RBD Binding Inhibition Assay
2
ACE2-RBD Interaction Blocking by LCB1
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The blocking activity of LCB1 peptide on the interaction of ACE2 and RBD proteins was also determined by flow cytometry. Briefly, an RBD protein at 2 μg/ml was mixed with serially diluted LCB1 and incubated at 4°C for 1 h. The mixture was then added to 5 × 105 of 293T/ACE2 cells and incubated at 4°C for 1 h. After being washed twice with PBS, the cells were incubated with 1:500 diluted Alexa Fluor 488-labeled rabbit anti-His tag antibody (Cell Signaling Technology, Danvers, MA, USA) at 4°C for 1 h. After two washes with PBS, cells were resuspended by FACS buffer and analyzed with a FACS CantoII instrument (Becton-Dickinson, Mountain View, CA, USA).
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