For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Loading of prestained molecular weight markers (#59329, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val62 of SARS-CoV-2 ORF8 protein.
The cause of the COVID-19 pandemic is a novel and highly pathogenic coronavirus, termed SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2). SARS-CoV-2 is a member of the Coronaviridae family of viruses (1). The genome of SARS-CoV-2 is relatively large and encodes up to 29 open reading frames (ORFs). These include ORF1a and ORF1b (further processed into 16 non-structural proteins), 9 accessory proteins, and 4 canonical structural proteins: spike (S), envelope (E), membrane (M), and nucleocapsid (N) (2). ORF8 is transcribed and translated from an ORF in the nested subgenomic RNA of the SARS-CoV-2 genome. It is one of the most divergent proteins in the SARS-CoV genome, and is believed to interfere with the immune response by binding to MHC-1 molecules and targeting them for lysosomal degradation via autophagy (3,4).
UniProt ID: P0DTC8
Entrez-Gene Id: 43740577
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