alexa 647 conjugated mouse anti sars cov 2 spike s1 subunit mab  (R&D Systems)

Journal: Scientific reports

Article Title: Effectiveness of VSV vectored SARS-CoV-2 spike when administered through intranasal, intramuscular or a combination of both.

doi: 10.1038/s41598-023-48397-7

Figure Lengend Snippet: Figure 1. VSV-S2 recombinant virus rescue amplification. (A) Illustrative genome for VSV- WT and VSV- S2. N: Nucleocapsid, P: Phosphoprotein, M: Matrix, G: Glycoprotein, L: Polymerase. To generate VSV-S2, Glycoprotein G is replaced with SARS-2 Spike Δ21. (B) Schematic illustration of VSV-S2 viral rescue and propagation. (C) Western blot for detection of viral proteins VSV-G, VSV-N, and SARS-CoV-2 spike S1. All lanes were run on the same gel and transferred to the same blot. The blot was subsequently cut into three sections immediately after protein transfer to membrane. Each section was then blocked with skim milk and detected with mouse anti-VSV nucleocapsid, mouse anti-VSV-G antibodies, or mouse anti-SARS-CoV-2 spike S1, respectively. Blots were imaged at an exposure of 10 s. Blots were cropped and resized for clarity and presentation. No other alteration of image was made. Original, uncropped and full size image of blots are shown in Supplementary Fig. S1. Plasmid expression of SARS-CoV-2 spike in 293 T was included as a positive control. Supernatants containing VSV-S2 or VSV-WT were harvested from Vero-TMPRSS2 cells (MOI = 0.1) 48 h post infection. Prior loading on SDS-PAGE gel, supernatants were filtered and centrifuged.

Article Snippet: Membrane blots were blocked in 5% skim milk/TBST (Tris-buffered saline with 0.1% Tween 20 (Cat# 194841, MP BioMedicals)) for 1 h at room temperature, followed by detection with primary antibodies mouse anti-S1 antibody (Cat# MAB105403, R&D systems), mouse anti VSV-N (Cat # MABF2348, Sigma Aldrich), or mouse anti-VSV-G (Cat #MABF2321, Sigma Aldrich).

Techniques: Recombinant, Virus, Amplification, Western Blot, Membrane, Plasmid Preparation, Expressing, Positive Control, Infection, SDS Page