Author: Bancroft, Tara; DeBuysscher, Blair L.; Weidle, Connor; Schwartz, Allison; Wall, Abigail; Gray, Matthew D.; Feng, Junli; Steach, Holly R.; Fitzpatrick, Kristin S.; Gewe, Mesfin M.; Skog, Patrick D.; Doyle-Cooper, Colleen; Ota, Takayuki; Strong, Roland K.; Nemazee, David; Pancera, Marie; Stamatatos, Leonidas; McGuire, Andrew T.; Taylor, Justin J.
Title: Detection and activation of HIV broadly neutralizing antibody precursor B cells using anti-idiotypes Document date: 2019_10_7
ID: 63yvpuqx_60
Snippet: The ZEUS IFA ANA HEp-2 Test System was used as recommended by the manufacturer (Zeus Scientific) with minor changes. Briefly, HEp-2-coated slides were incubated with 25 µl antibody at 0.1 mg/ml for 30 min at room temperature. Slides were then washed three times in 1× DPBS followed by the incubation of 25 µl of 0.01 mg/ml goat anti-human IgG Alexa Fluor 594 (Thermo Fisher Scientific) in 1× DPBS for 30 min at room temperature in the dark. After.....
Document: The ZEUS IFA ANA HEp-2 Test System was used as recommended by the manufacturer (Zeus Scientific) with minor changes. Briefly, HEp-2-coated slides were incubated with 25 µl antibody at 0.1 mg/ml for 30 min at room temperature. Slides were then washed three times in 1× DPBS followed by the incubation of 25 µl of 0.01 mg/ml goat anti-human IgG Alexa Fluor 594 (Thermo Fisher Scientific) in 1× DPBS for 30 min at room temperature in the dark. After washing three times with 1× DPBS, slides incubated with ZEUS IFA ANA HEp-2 Test System fixative, and coverslips were applied. Images were acquired using the EVOS Cell Imaging System (Thermo Fisher Scientific) and analyzed using ImageJ to determine the average Alexa Fluor 594 fluorescence per HEp-2 cell for each image.
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