Do you want to analyze Hbs or the rest of proteins in RBCs?

If you want to analyze Hbs this should help:

Sample preparation for protein electrophoresis

Blood samples were collected in heparinized tubes and red blood cells (RBCs) were separated from plasma by centrifugation at 3,000 g for 1 minute. For the protein analysis of hemoglobin, 100 ul of red blood cells (RBCs) were washed twice with two volumes of ice-cold saline solution (1 mM Tris, pH 8.0, 200 mM NaCl) and collected by centrifugation at 3,000 × g for 5 min. at 4°C. The RBCs were lysed in 300 ul of cold buffer (1 mM Tris, pH 8.0, 1 mM phenylmethylsulphonyl fluoride). To complete hemolysis, samples were placed on ice for 10 minutes and then at -80°C for 15 min. Cellular debris was pelleted by centrifugation at 15,000 × g for 15 minutes at 4°C and the hemolysate was stored in 100 mM NaCl at -80°C. Protein concentration of the hemolysate was measured using the Bradford method [52].

From Borza et al, BMC Genetics 2009, 10:51 Open access

It is possible. Hemoglobin is a protein. Separating it from a full RBC extract will be easiest by size exclusion chromatography, SDS-PAGE or anion exchange. Let me know if I can help you.

Human RBCs or from other vertebrates? For all mammalian RBC just use a hypotonic solution and all Hb will be released. This is a classic method to obtain the cytoskeleton and membrane proteins.

Thanks Maria Can you tell in brief how to separate Hb by SDS PAGE

Presently I am using Hypotonic solution but it takes 15 - 20 wash to completely remove Hb Is their any specific haemolysis buffer and sometimes proteins seems to be aggregate and how to dissolve it completely. kindly suggest me.

Do you want to analyze Hbs or the rest of proteins in RBCs?

If you want to analyze Hbs this should help:

Sample preparation for protein electrophoresis

Blood samples were collected in heparinized tubes and red blood cells (RBCs) were separated from plasma by centrifugation at 3,000 g for 1 minute. For the protein analysis of hemoglobin, 100 ul of red blood cells (RBCs) were washed twice with two volumes of ice-cold saline solution (1 mM Tris, pH 8.0, 200 mM NaCl) and collected by centrifugation at 3,000 × g for 5 min. at 4°C. The RBCs were lysed in 300 ul of cold buffer (1 mM Tris, pH 8.0, 1 mM phenylmethylsulphonyl fluoride). To complete hemolysis, samples were placed on ice for 10 minutes and then at -80°C for 15 min. Cellular debris was pelleted by centrifugation at 15,000 × g for 15 minutes at 4°C and the hemolysate was stored in 100 mM NaCl at -80°C. Protein concentration of the hemolysate was measured using the Bradford method [52].

From Borza et al, BMC Genetics 2009, 10:51 Open access

If you are doing small scale removal of hemoglobin you could try using haptoglobin sepharose.

http://jasn.asnjournals.org/content/13/2/423.full.pdf+html

Thanks Tudor I will try this protocol and how to dissolve protein completely sometimes protein found to be Fibrous

Dear Tudor Borza,

can i use 50mM ammonium bicarbonate buffer or any other Mass spect compatible detergents/buffers to dissolve / store hemolysate to store insteade of 100 mM NaCl; because i have to do mass spect analysis for the hemoglobin.

Please give me your valuable suggestion..

Many years ago Seman M. .et al. (Eur. J. Immunol., 1971, 2, 387) have published a method of  isolation of water soluble proteins from sheep red blood cells. Perhaps  some details of this procedure  may.be useful for your. Really it is very difficult to remove Hb. Are you sure that it is necessary for your purpose?