HemoVoid™ Applications

Cooperating G6PD mutations associated with severe neonatal hyperbilirubinemia and cholestasis.

The paper titled, “Cooperating G6PD mutations associated with severe neonatal hyperbilirubinemia and cholestasis” uses HemoVoid™ for performing native gel electrophoresis and immunoblotting on blood samples from the patient and control subjects that were lysed and depleted of hemoglobin. Subsequent to using HemoVoid™, non-bound protein was eluted at pH 9.8 and placed in native sample buffer, pH 6.8 and analyzed by native gel electrophoresis in polyacrylamide gradient gels of 4–15%. Separating hemoglobin from erythrocytes by contacting erythrocytes with a hypotonic buffer solution at a rate sufficient to render the release of hemoglobin from said erythrocytes without significant lysis. The hemoglobin is then separated from the erythrocytes. Hemovoid allows for the purification of hemoglobin solutions of DNA, endotoxins and phospholipids by contacting the hemoglobin solutions with an anion exchange medium.

Functional 20S proteasomes in mature human red blood cells

Hemovoid is used to study the functional 20S and/or 26S proteasomes within red blood cells (RBCs; depleted of reticulocytes and leukocytes).Using methods such as double-immunofluorescence confocal microscopy to localize mature RBCs, proteasomes are isolated from mature RBC. Aftering using Hemovoid, a two-dimensional differential in-gel electrophoresis (2D-DIGE)approach was used to determine if proteasome-dependent protein degradation occurs within mature RBCs.