Mice were immunized subcutaneously in the flanks on day zero with 50 μg NP-KLH (Biosearch Technologies) emulsified in 100 μl Imject Alum (ThermoScientific). Serum and draining inguinal lymph nodes for flow cytometric analysis were collected on day 12 post-immunization. Affinity assays were performed as described previously29 (link). Briefly, plates (Corning, New York, NY) were coated at with 10 μg/ml of either NP-27-BSA (total) or NP-2-BSA (high-affinity) per well (Biosearch Technologies). Antibody titers are given as –log2 dilution × 40. Positive titers were defined as three standard deviations (SD) above the mean values of the negative controls41 (link).
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NP-KLH Immunization and Antibody Affinity Analysis
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Alum
Antibody
Assays
Dilution
Flow cytometric
Immunization
Inguinal
Lymph nodes
Mice
Np 27
Np klh
Serum
Corresponding Organization :
Other organizations : Kantonsspital St. Gallen, University Hospital of Zurich, University of Zurich, Osaka University, Babraham Institute
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Variable analysis
independent variables
- Immunization with 50 μg NP-KLH emulsified in 100 μl Imject Alum on day zero
- Serum and draining inguinal lymph nodes for flow cytometric analysis collected on day 12 post-immunization
- Antibody titers measured as -log2 dilution × 40
- Not explicitly mentioned
- Positive control: Not specified
- Negative control: Mean values of negative controls used to define positive titers as three standard deviations above the mean
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