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Fig. 2.
Selective recognition of human growth hormone. The sample, consisting of
human growth hormone (GH) and human serum albumin (HSA), was applied (a) directly
on an anion exchanger for analysis, (b) on a blank column prepared in the absence of
growth hormone and the eluated protein fraction was analyzed on the same ion-exchanger,
and (c) on a column prepared in the presence of growth hormone and the eluated protein
fraction was analyzed on the ion-exchanger. A comparison of the three chromatograms
shows that the growth hormone was adsorbed only onto the gel synthesized for specific
recognition of this protein (i.e. the gel prepared in the presence of the growth hormone).
lysozyme (5 mg) were dissolved in 2.94 mL of water. Following the addi-
tion of 30
L of a 10% (w/v) aqueous solution of ammonium persulfate
and deaeration by aspiration, the mixture was supplemented with 30
µ
L
of a 5% (v/v) aqueous solution of TEMED. The polymerization was
allowed to proceed for three hours. The gel, which had the composition
T6, C5 (Hjertén, 1962), was granulated by pressing it through a 100-mesh
net and packed into a 5
µ
L plastic Eppendorf syringe (with the piston
removed) that had been fitted at the outlet with a piece of a cut nylon
stocking. The column was washed for three hours with 10 mL of a 10%
(w/v) SDS solution in 10% (v/v) acetic acid to desorb the three proteins,
but was still somewhat colored, indicating that hemoglobin and
cytochrome
c
(and therefore probably also lysozyme) could not
be released completely. The column was equilibrated with 12 mL of
100 mM sodium phosphate, pH 6.2, for one hour and was then stored for
two days before it was tested for adsorption of hemoglobin, cytochrome
c
and lysozyme by applying 35
µ
L of these proteins dissolved in 0.1 M
sodium phosphate, pH 6.2, at a final concentration of 0.020%, 0.085% and
0.085% (w/v), respectively. The column was washed with 100 mM
µ
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