Neutralization assays were performed by incubating Abs or sera with HIV-1 pseudoviruses that are capable of one round of infection to TZM-BL cells (Walker et al., 2009 (link)). Serum depletion was performed with Tosylactivated Dynabeads (Life Technologies) conjugated with BSA or with WT and N332A 92BR020 rgp120 proteins. Serum samples were depleted of Abs binding to these proteins through multiple rounds of immunoprecipitation (Li et al., 2009 (link)) as confirmed by ELISA, prior to use in neutralization assays.
Tosylactivated dynabeads
Manufactured by Thermo Fisher Scientific
Sourced in United States
Tosylactivated Dynabeads are magnetic beads designed for covalent coupling of proteins, peptides, oligonucleotides, and other ligands. The beads have a tosyl-activated surface that allows for simple and efficient immobilization of a wide range of molecules. Tosylactivated Dynabeads provide a versatile platform for various applications in biotechnology and research.
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Lab products found in correlation
3 protocols using tosylactivated dynabeads
1
HIV-1 Neutralization Assay with Serum Depletion
2
Affinity Purification of Serum Proteins
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Five milligrams of tosylactivated dynabeads (Life Technologies, Carlsbad, CA, USA) were either conjugated to 100 μg mAb RETC-2, mAb RETC-3, or the respective IgG isotype controls according to the manufacturer’s protocol. Pooled normal human serum (NHS, 1.5 mL) was incubated with mAb-coupled dynabeads (50 μL, 1 h). After washing, proteins were eluted using non-reducing Laemmli sample buffer (10 min, 95°C) (26 (link)).
3
Antibody-Based Protein Isolation from Serum
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Tosylactivated dynabeads (Life Technologies, Carlsbad, USA, 5 mg) were conjugated to 100 µg mAb 1340 according to the manufacture's protocol. Pooled human Serum (NHS, 1.5 mL) was incubated with mAb 1340-dynabeads (50 µL, 1 h). After washing with wash buffer, proteins were eluted using non-reducing Laemmli sample buffer and denaturation at 95°C (10 min) [41] .
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