HemoVoid™ Allows Researchers to Study G6PD Mutations Associated With Severe Neonatal Hyperbilirubinemia and Cholestasis
HemoVoid™ Allows Researchers to Study G6PD Mutations Associated With Severe Neonatal Hyperbilirubinemia and Cholestasis
A new report published in the journal Pediatric Blood & Cancer by Mizukawa, B.et al features the use of HemoVoid™™ on erythrocyte samples and rabbit anti-human G6PD polyclonal antibody. Scientists are able to perform genetic analysis to gather information deciphering the association between mutations and their predisposition to disease. Researchers from Department of Hematology-Oncology and Gastroenterology at the Children’s Hospital Medical Center- University of Cincinnati College of Medicine studied erythrocytes to identify causative mutations in sporadic cases to find novel causative mutation for neonatal hyperbilirubinemia.
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.
Glucose-6-phosphate dehydrogenase deficiency is an X-linked inherited disorder that is the most common enzyme deficiency worldwide. If you do not remove the erythrocyte it is difficult to study diseases which involve enzyme deficiencies. Biotech Support Group has two types of hemoglobin removal agents. HemogloBind™ is engineered for a high degree of hemoglobin binding selectivity and does not cross react with most common serum components. HemogloBind™'s binding capacity for hemoglobin is approximately 10mg/ml of hemoglobin. HemogloBind™ is used mostly on hemolysed serum. HemoVoid™ does not bind to hemoglobin but does bind to other proteins. So it removes hemoglobin from erythrocyte lysate samples while concentrating low abundance, and/or low molecular weight proteins. HemoVoid™ is used mostly on red cell lysates for proteomic studies of the preserved enzymes that are retained in elution fractions.
References
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