30G 



BIOCHEMICAL GENETICS 



but the minor components of multiple hemoglobins become more readily visible 

 following staining (figure 41) with a saturated solution of Amido black 1 OB in methanol, 

 H 2 0, and acetic acid (5:5:1). It is more difficult to estimate the quantity of different 

 fractions of hemoglobin in starch-gels than on paper strips. Photoelectric devices 

 have been used with preparations of starch gels made temporarily transparent by boiling 

 them for 30 seconds in a 10 per cent solution of acetic acid. 1342 Permanent preparations 



Fig. 41. Comparison of murine oxyhemoglobins analyzed by starch-gel 



ELECTROPHORESIS. 



B 



Red-cell lysates were electrophoresed for four hours; borate buffer; ionic strength 

 0.025, pH 8.5, 6 volts/cm., 6 milliamps. Hemoglobin was stained with Amido black 10B. 

 Above (A). Single hemoglobin of strain SeC mice. 

 Below (B). Diffuse hemoglobin of strain BALB/c mice. 



for photometric analyses can be made 361 by mounting stained, transparent starch-gel 

 blocks in agar. Quantitative analysis of eluted hemoglobin can also be made after 

 freezing the gel, which makes the gel spongelike, and removing the pigment by applying 

 pressure. 



The electrophoretic mobility of proteins is not identical in starch gel and on 

 paper, 1020 presumably due to greater adsorption of proteins on paper. Although two- 

 dimensional electrophoresis — paper in one direction and starch gel in another at right 

 angles to it — can be used for some protein separations, this system is perhaps not useful 

 for hemoglobin. However, because of the greater resolution two-dimensional electro- 



