You could use centrifugation plus washes or gel filtration.

Hi Indrajit

How have you lysed erythrocytes?

If you have used the hypotonic lysis method recommended for the preparation of erythrocyte membrane ghosts, then it is a matter of repeated washes and centrifugation of the ghosts until you obtain Hb-free white ghosts - as Rudy suggested.  If you used old blood to prepare the lysates it will be harder to obtain Hb-free ghosts as some of the denatured Hb will sediment with the ghosts.

Regards, Rosemary

Need to wash the cells to remove plasma protein. After lysate filtration using 0,45 uL filter you can obtain hemoglob

Use hypotonic medium which as water to lyse the red cells. Then centrifuge at high speed. Filter the supernatant and take the filterate as your Hb solution. Add more water to the debris mixed, spin at high speed and filter the supernatant and add the filterate the precious filterate Hb solution.