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Merck

A Protocol for Studying HIV-1 Envelope Glycoprotein Function.

STAR protocols (2020-12-31)
Sneha Ratnapriya, Angela Chov, Alon Herschhorn
ABSTRACT

HIV-1 envelope glycoproteins (Envs) bind to CD4 receptor and CCR5/CXCR4 coreceptor and mediate viral entry (Feng et al., 1996; Herschhorn et al., 2016, 2017; Kwong et al., 1998). HIV-1 Envs are the sole target of neutralizing antibodies and a main focus of vaccine development (Flemming et al., 2018). Here, we provide a step-by-step protocol to measure Env sensitivity to ligands, cold, and small molecules, as well as to study viral infectivity and to dissect parameters affecting HIV-1 Env function. For complete details on the use and execution of this protocol, please refer to Harris et al. (2020).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
trans-1,2-Diaminocyclohexane-N,N,N′,N′-tetraacetic acid monohydrate, ACS reagent, for complexometry, 98%
Sigma-Aldrich
DL-Dithiothreitol solution, BioUltra, for molecular biology, ~1 M in H2O