Anti-gp120 Ab titers were measured by D7324-capture ELISA as described previously (35 (link),44 (link),47 (link)). Reactivity of Env proteins with MAbs or receptor mimics were measured by anti-trimer ELISA as described previously using Ni-NTA HisSorb 96-well plates (Qiagen, Venlo, The Netherlands)(10 (link),34 (link)–36 (link),47 (link)). Equal input levels of proteins were verified by SDS-PAGE followed by western blot analysis. All ELISA data are representative of at least three independent experiments, each using Env proteins derived from independent transfections. Endpoint titers were calculated using GraphPad Prism (version 5.03) by determining the serum dilution at which the optical density (OD450) was three times above the background OD450 signal (without animal sera).
Protocol detail
Anti-gp120 Antibody Titration by ELISA
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Animal
Dilution
Elisa
Env proteins
Gp120
Mabs
Optical
Prism
Proteins
Sds page
Sera
Transfections
Western blot analysis
Corresponding Organization :
Other organizations : Academic Medical Center, University of Amsterdam, Cornell University
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Variable analysis
independent variables
- Anti-gp120 Ab titers measured by D7324-capture ELISA
- Reactivity of Env proteins with MAbs or receptor mimics measured by anti-trimer ELISA
- Optical density (OD450) of ELISA assays
- Equal input levels of proteins verified by SDS-PAGE followed by western blot analysis
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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