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Antigen retrieval was performed in a microwave for 10 min using the appropriate antigen retrieval buffer for each antigen (Table 2).
The following antibodies were raised and affinity purified using the appropriate antigen by the DSTT: sheep anti-FAF1 (S370D; antigen human FAF1, 1-650), rabbit anti-VAPB (R2986 and R2986; andigen human VAPB, 1-210).
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CTL clones were established from PBMCs or CTL lines by either limiting dilution or by using the appropriate antigen-specific tetramers in combination with MACS anti-PE-microbeads (Miltenyi Biotec).
Antibodies were affinity purified from immune sera using the appropriate antigens.
Following conjugation, the reactivity of the MAb porphyrin conjugates was compared to that of the unconjugated MAb using the appropriate antigen-positive cell lines by flow cytometry.
Antigens were visualized using the appropriate fluorophore-conjugated secondary antibodies (Alexa Fluor ® 555 goat anti-mouse IgG (H+L), 1 500, Life Technologies Alexa Fluorr ® 488 goat anti-rat IgG, 1 500, Life Technologies).
Using the appropriate chip-specific dilution buffers, we diluted the serum samples 1 10 for use in the synovial antigen 1 and 2, CID 3, and CID 4 chips, and 1 100 for use in the CID 1 chips.
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The labelled cells are then used to detect the appropriate antigen or antibody in a simple agglutination test.
In general, better performance in serological screening depends on the use of the appropriate antigens and adjustment of cut-off values [ 22].
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