Centrifuge samples at 1200g for five minutes.

It diluted 20μl of the packed red

cells with 150μl of the haemolysing chemical reagent. It combined gently and
leave for a minimum of five minutes. If clarified haemolysates are used, dilute
40μl of 100g/l haemolysate with 150μl of lysing chemical reagent.
Disconnected the power supply, prepare the electrophoresis tank by putting
equal amounts of TEB buffer in every of the outer buffer compartments. Wet 2
chamber wicks within the buffer, and place one on every divider/bridge support
making sure that they develop best contact with the buffer.
Soak the cellulose acetate by lowering it slowly into a reservoir of buffer. Leave
the cellulose acetate to soak for a minimum of five minutes before use.
Fill the sample well plate with 5μl of every diluted sample. Manage and conceal
with a 50nm coverslip or a ‘short’ glass to impede evaporation. Load a second
sample well plate with Zip Zone Prep solution.
Clean the device tips instantly before use by loading with Zip Zone Prep Solution
then applying them to a blotter.
Remove the cellulose acetate strip from the buffer and blot double between 2
layers of neat and clean blotting paper. Do not permit the cellulose acetate to dry.
Load the device by depressing the tips into the sample wells two time and apply
this 1st loading onto some clean blotting paper. Reload the device and apply the
samples to the cellulose acetate.
Place the cellulose acetate plates across the bridges, with the plastic side
topmost. Place 2 glass slides across the strip to manage of best contact.
Electrophorese at 350V for twenty five minutes.
After twenty five minutes electrophoresis, right away transfer the cellulose
acetate to Ponceau S and fix and stain for five minutes.
Remove excess stain by cleaning for five minutes within the 1st ethanoic
acid reservoir and then for ten minutes in every of the remaining 2 reservoir. Blot
once, through clear blotting paper and leave to dry.
Label the membranes and save through a plastic envelope for protection.