ELISA for Recombinant NTS-DBL1α Antibodies
Corresponding Organization :
Other organizations : Karolinska Institutet
Variable analysis
- Concentration of immune sera samples
- Optical density (OD) at 405 nm, as a measure of IgG antibody levels against the recombinant NTS-DBL1α antigen
- Coating of Maxisorp plates with 1 μg of the recombinant NTS-DBL1α antigen dissolved in 15 mM Na2CO3 and 35 mM NaHCO3 (pH 9.6)
- Washing of plates three times with PBS containing 0.1% Tween 20 (PBST)
- Blocking of plates with 1% bovine serum albumin (BSA) in PBST for 1 h at room temperature
- Washing of plates three times with PBST before adding samples
- Incubation of samples in triplicate for 1 h at room temperature
- Washing of plates three times with PBST after sample incubation
- Incubation of plates with alkaline phosphatase-conjugated goat anti-human IgG (1:1000 in PBS) for 1 h at room temperature
- Washing of plates three times with PBST before development
- Development of plates with SigmaFast p-nitrophenyl phosphate tablets for 15 min
- Positive control: A control pool of six non-malaria exposed Swedish donors
- Negative control: Control wells without serum (background)
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