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The deep red coloured 30 nm colloidal gold nano-particles was prepared by trisodium citrate method. The colloidal gold particles showed a peak at 527 nm using spectrophotometric scanning between 400 to 700 nm and particle size of 30 nm was further confirmed in transmission electron microscope (data not shown), which provided the best basis for preparation of the LFIA. To find out the required concentration of IgG for getting stable colloidal gold conjugates, we tested colloidal gold particles with different concentration of IgG and variable pH. The results showed that 16 µg/ml of IgG concentration coupled with pH 9.0 was optimized for stable conjugation of IgG.

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