Plaque areas were measured in CECs exactly as previously described32 (link) using anti-MDV chicken sera and goat anti-chicken IgY-Alexa Fluor 488 secondary antibody (Molecular Probes, Eugene, OR). Digital images of 36 individual plaques were collected using Nikon Eclipse-Ti-E inverted fluorescent microscope and plaque areas were measured using ImageJ33 version 1.41o software. Whisker plots were generated using Microsoft Excel 365 and significant differences were determined using IBM SPSS Statistics Version 28 software Package (https://www.ibm.com/analytics/spss-statistics-software ).
Goat anti chicken igy alexa fluor 488 secondary antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
Goat anti-chicken IgY-Alexa Fluor 488 secondary antibody is a fluorescently labeled antibody used for the detection and quantification of chicken immunoglobulin Y (IgY) in various research and diagnostic applications.
Automatically generated - may contain errors
Lab products found in correlation
Evos fl cell imaging system, by Thermo Fisher Scientific (3 mentions)
Eclipse ti e inverted fluorescent microscope, by Nikon (1 mentions)
Spss statistics version 27, by IBM (1 mentions)
Anti mouse ig alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Mouse anti flag m2, by Merck Group (1 mentions)
4 protocols using goat anti chicken igy alexa fluor 488 secondary antibody
1
Measuring Plaque Areas in CECs
2
Quantification of Viral Plaque Sizes
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Plaque areas were measured in CEC cultures exactly as previously described49 (link) using anti-GaHV-3 chicken sera and goat anti-chicken IgY-Alexa Fluor 488 secondary antibody (Molecular Probes, Eugene, OR). Digital images of 50 individual plaques were collected using an EVOS FL Cell Imaging System (Thermo Fisher Scientific) and compiled with Adobe Photoshop version 21.0.1. Plaque areas were measured using ImageJ50 version 1.53d software (http://imagej.nih.gov/ij ). Box and Whisker plots were generated, and significant differences were determined using IBM SPSS Statistics Version 27 software.
3
Visualizing Reconstituted Virus Plaques
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CECs were infected with reconstituted viruses in 6-well tissue culture plates at 100 plaque-forming units (PFU) per well. At 5 days post-infection (dpi), cells were fixed with PFA buffer (2% paraformaldehyde, 0.1% Triton X-100) for 15 min and then washed twice with PBS. The plaques were fluorescent red and green but for double confirmation the fixed cells also were stained with anti-MDV chicken sera plus goat anti-chicken IgY-Alexa Fluor 488 secondary antibody (Molecular Probes, Eugene, OR, USA). The virus plaques were observed using an EVOS FL Cell Imaging System (Thermo Fisher Scientific, Waltham, MA, USA) and compiled using Adobe Photoshop version 21.0.1.
4
Immunofluorescence Assay for Virus Plaque Detection
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CEC cultures were infected with different r301B/1 viruses on sterile glass coverslips at 100 plaque-forming units (PFU) per well. At 5 days post-infection (p.i.), cells were fixed with PFA buffer (2% paraformaldehyde, 0.1% Triton X-100) for 15 min and then washed twice with PBS. Fixed coverslips were blocked in 10% neonatal calf serum and stained with anti-GaHV-3 chicken sera and goat anti-chicken IgY-Alexa Fluor 488 secondary antibody (Molecular Probes, Eugene, OR). To detect 3 × Flag301BgC, mouse anti-Flag M2 (Sigma-Aldrich) was used. Anti-gC monoclonal A6 (kindly provided by Jean-Francois Vautherot, INRA, Nouzilly, France) antibody48 (link) was used to detected MDV gC expression. Anti-mouse Ig Alexa Fluor 488 (Molecular Probes, Eugene, OR) was used as secondary antibody for both anti-Flag and -MDV gC monoclonal antibodies. The virus plaques were observed using an EVOS FL Cell Imaging System (Thermo Fisher Scientific) and compiled using Adobe Photoshop version 21.0.1.
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