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Novel monoclonal antibodies to the SERINC5 HIV-1 restriction factor detect endogenous and virion associated SERINC5

MAbs. 2020-08; 
Sebastian Molnar, Lindsay Wieczorek, Michelle Zemil , Bianca Schulte, Elizabeth Martinez , Syna Gift, Lan Tang, Hendrik Streeck, Robert A Gramzinski, Nelson L Michael, Gordon Joyce , Victoria R Polonis
Products/Services Used Details Operation
Codon Optimization The SERINC5 transcript variant 1 (GenBank accession numberNM_001174072.3) was codon-optimized with the GeneSmartR codon optimization tool (GenScript, Piscataway, NJ) and the gene was cloned in CMVR-3c-His using XbaI and BamHI restriction sites...We used the OptimunAntigenTM tool (GenScript, Piscataway, NJ)30 to design maximally immunogenic peptides and ensure that the three peptides regions did not overlap and had the best immunogenicity surface exposure and hydrophilicity scores possible (Table 1)... Plates were washed four times with PBST, and then a horse radish peroxidase-conjugated anti-mouse secondary antibody(GenScript Cat# A00160) was added and incubated for 30 minutes at 37°C. Get A Quote
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摘要

SERINC5 is a multi-pass transmembrane protein that is thought to play a role in serine incorporation during cellular membrane biosynthesis. This protein has also been identified as a human immunodeficiency virus Type 1 (HIV-1) restriction factor. The paucity of monoclonal antibodies (mAbs) against SERINC5 has posed a challenge for the study of the endogenous protein. Here we report the development of novel anti-SERINC5 mAbs that target three distinct loops on the protein. We demonstrate that these SERINC5 mAbs can be used to detect endogenously expressed SERINC5 protein in various cell lines using Western blot, whole-cell ELISA, flow cytometry, and immunocytochemistry. We further show that some of these antibod... More

關鍵詞

HIV-1 restriction factor; SERINC5; immunocytochemistry; monoclonal antibody; multi-pass transmembrane protein; serine incorporator; virus capture.
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