Nunc Maxsorp ELISA plates were coated overnight with 5 μg/ml dsDNA from Lambda bacteriophage (New England BioLabs) using Reacti-Bind DNA Coating Solution (Thermo Scientific) according to the manufacturer’s instructions. After blocking with PBS/2% BSA, sera were diluted with PBS/1% BSA and added at various concentrations. Plates were washed with PBS /0.05% Tween20 and bound antibody was detected using pooled biotin labelled anti IgGs listed above prior to Streptavidin-HRP addition (BD Biosciences). Anti RNA antibodies were determined as above, only Baker’s Yeast RNA (Sigma) was used to coat the plate.
Lambda bacteriophage
Manufactured by New England Biolabs
Lambda bacteriophage is a virus that infects Escherichia coli bacteria. It is a temperate phage, meaning it can either undergo lytic or lysogenic life cycles. The core function of lambda bacteriophage is to infect and replicate within the host E. coli cell.
Automatically generated - may contain errors
2 protocols using lambda bacteriophage
1
ELISA for dsDNA and RNA Antibodies
2
Enzyme-Linked Immunosorbent Assay for Anti-DNA Antibodies
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Nunc MaxiSorp ELISA plates were coated overnight with 5 μg/ml dsDNA from Lambda bacteriophage (New England BioLabs) using Reacti-Bind DNA Coating Solution (Thermo Scientific) according to the manufacturer’s instructions. After blocking with PBS/2% BSA, sera were diluted with PBS/1% BSA and added at various concentrations. Plates were washed with PBS/0.05% Tween 20, and bound Ab was detected using pooled biotin-labeled anti-IgGs listed earlier before Streptavidin-HRP addition (BD Biosciences). Anti-RNA Abs were determined as described earlier; only Baker’s Yeast RNA (Sigma) was used to coat the plate.
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