Immunoblotting of Viral Antigen Proteins

Antigen preparations were resolved by SDS-PAGE, transferred to PVDF membranes (EMD Millipore), and probed with antibodies as published [28] (link). In some experiments, antigens resolved by SDS-PAGE were stained with SimplyBlue™ SafeStain (ThermoFisher Scientific). The mouse antibody His-Tag (EMD Millipore) was used in experiments with His-Tag BatA protein. Murine antibodies against a peptide encompassing BatA residues 208–221 (GenScript custom polyclonal antibody production services) were used in experiments with PIV5-BatA virus. Goat anti-mouse antibodies conjugated to HRP (SouthernBiotech) were utilized for detection.

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Lafontaine E.R., Chen Z., Huertas-Diaz M.C., Dyke J.S., Jelesijevic T.P., Michel F., Hogan R.J, & He B. (2018). The autotransporter protein BatA is a protective antigen against lethal aerosol infection with Burkholderia mallei and Burkholderia pseudomallei. Vaccine: X, 1, 100002.

Publication 2018

PVDF membranes SimplyBlue™ SafeStain His-Tag

Anti antibodies Antibodies Antibody Antibody production Antigens Goat Membranes Mouse Murine Peptide Protein Pvdf Sds page Virus

Corresponding Organization : University of Georgia

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Variable analysis

independent variables

Antigen preparations resolved by SDS-PAGE
Antigens resolved by SDS-PAGE and stained with SimplyBlue™ SafeStain

dependent variables

Antigen preparations transferred to PVDF membranes
Antigens probed with antibodies

control variables

Experiments with His-tag BatA protein
Experiments with PIV5-BatA virus

controls

Mouse antibody His-Tag used in experiments with His-tag BatA protein
Murine antibodies against a peptide encompassing BatA residues 208–221 used in experiments with PIV5-BatA virus
Goat anti-mouse antibodies conjugated to HRP used for detection

Annotations

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