UC San Diego Human Research Protections Program (irb.ucsd.edu) approved IBC Protocol 20004 for the use of SARS-CoV-2 convalescent plasma. Anonymized plasma samples of SARS-CoV-2 convalescent individuals (N = 19) were obtained from UCSD. The patients gave informed consent. Individuals were confirmed to be infected in the previous 3–10 weeks by PCR and lateral flow assay. All individuals were symptomatic with mild to moderate-severe symptoms. Serum samples (DS-626-G and DS-626-N, Seracare) purchased before SARS-CoV-2 pandemic were used as a negative control. SARS-CoV-2-specific binding antibodies in plasma samples were measured as described above. Cross-adsorbed goat anti-human IgG (H + L) secondary antibody (A18811, Invitrogen) was used at a dilution of 1:3000.
Goat anti human igg h l secondary antibody
Manufactured by Thermo Fisher Scientific
Goat anti-human IgG (H + L) secondary antibody is a laboratory reagent used to detect the presence of human immunoglobulin G (IgG) in biological samples. It is a secondary antibody that binds to the heavy and light chains of human IgG, allowing for the identification and quantification of this antibody in various immunoassays and research applications.
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2 protocols using goat anti human igg h l secondary antibody
1
Characterization of SARS-CoV-2 Convalescent Plasma
2
Quantifying TF-TCB Binding to CD3 and TF
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The binding ability of TF-TCB to cell surface CD3 and TF was measured by flow cytometry. MDA-MB-231 cells (2 × 105 cells/well) or Jurkat cells (2 × 105 cells/well) were placed into 96-well U bottom-plate (Corning, Corning, NY, USA) after resuspension in FACS buffer (2% fetal bovine serum in PBS buffer). Test antibodies were serially diluted 1:3 in FACS buffer and added to the cells in a total volume of 100 μL. After incubation on ice for 30 min, cells were washed 3 times with FACS buffer and resuspended in 100 μL 1:200 goat anti-human IgG (H + L) secondary antibody (FITC-labeled, Invitrogen) in FACS buffer. Cells were incubated for another 30 min on ice. After washing 3 times with FACS buffer, mean fluorescence intensity (MFI) of cells was analyzed on FACSCalibur (Beckman Coulter, Fullerton, CA, USA). The binding EC50 value based on MFI was calculated in GraphPad Prism using non-linear regression analysis for single site binding26 (link).
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