By P. Alexander, R. J. Block
Read Online or Download Determination of the Size and Shape of Protein Molecules. A Laboratory Manual of Analytical Methods of Protein Chemistry (Including Polypeptides): Volume 3 PDF
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Extra info for Determination of the Size and Shape of Protein Molecules. A Laboratory Manual of Analytical Methods of Protein Chemistry (Including Polypeptides): Volume 3
Sample text
Q" is correlated with the permeability of the membrane by proteins. For a membrane of uniform permeability 8 cm long, 1-13 cm diameter and 2 28-4 c m surface area, filled with water and subjected to a pressure equivalent to 600 cm water, the values for q" convenient for investigation of proteins of different particle sizes are as follows: q" = 0*01, proteins of molecular weight 6000 are retained; q" = 0-03, haemoglobin and myoglobin are retained; q" = 0*1, haemoglobin is retained but the membrane may be stained and the permeability of the membrane and the concentration of haemoglobin must thereby be reduced; q" = 0-2, serum proteins are retained, haemoglobin may diffuse through slowly; q" = 0-5, haemocyanin is retained, haemoglobin diffuses through the membrane.
Before a membrane is mounted on the rubber tubing at present available it may be necessary to taper the rubber on a small grinding wheel, used solely for this purpose. Larger membranes, useful for preparative dialysis, may be mounted on rubber stoppers bored with one hole, through which a glass tube of about 6 cm length has been inserted. After mounting, membranes should be tested for leaks and for permeability. A piece of glass tube is sealed to each end of a graduated tube, conveniently made from a discarded 25 or 50 ml burette and the rubber m o u n t of the membrane is fixed to one end of the graduated tube.
After mounting, membranes should be tested for leaks and for permeability. A piece of glass tube is sealed to each end of a graduated tube, conveniently made from a discarded 25 or 50 ml burette and the rubber m o u n t of the membrane is fixed to one end of the graduated tube. Membrane and graduated tube are filled with water and the other end of the tube is connected to a glass bulb filled with air which can be subjected to pressures u p to 700 m m Hg. Pressure forces droplets of water, which should be of approximately uniform size, through the membrane.