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prepare platelets from control and immunized mice distribute 2.5 x106 platelets per well on F-bottomed MaxiSorp Nunc-Immuno microplates (GibcoBRL, Roskilde, Denmark), centrifuge at 3000 rpm, 7 min, 10ѓC. fix with PBS-PF
, incubate for 30 min at 37°C. ( 5g powdered milk in 95 ml PBS)5 Rinse with PBS 3x.6 Add 50 µl of antibody, incubate 30 min at 37°C.7 Rinse with PBS 3x.8 Add 50 µl of goat anti-mouse IgG conjugate with alkaline phosphatase.(1 µl of conjugate in 4 ml of PBS
Mapping Protein Distributions on Polytene Chromosomes by Immunostaining
To immunostain using enzyme-coupled secondary antibodies: i. Rinse slides in PBS and remove excess solution. ii. Add 40 µL of secondary antibody (e.g., anti-rabbit IgG [Fc] HRP conjugate, 1:100 dilution
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