Fig. 2.

Comparison of the amount of Hp and Angiotensin II present in 2 kDa dialysis bags. 1 ml Hp a or angiotensin II b solution (0.1 or 0.5 mg/ml) was placed in a dialysis bag of 2 kDa cutoff (Slide-A-Lyzer Dialysis Cassettes, Pierce) and subjected to dialysis against PBS for 24 h. Aliquots (20 μl) of the 24 h dialysate were subjected to HPLC analysis. The amount of Hp or angiotensin II in the bag before and after dialysis was calculated from the area under the curve and from a standard curved obtained by HPLC analysis of Hp or angiotensin II (0-0.5 mg/ml). There was <5% peptide associated with the dialysis cassette itself. a The area under the curve for Hp (0.5 mg/ml) before dialysis was 15758985 and after 24 h dialysis was 7014526. b The area under the curve for angiotensin II (0.5 mg/ml) before dialysis was 10389021 and after 24 h dialysis was 1490275. Insets in a and b show the amount of hemopressin or angiotensin II peptide in the dialysis bag before and 24 h after dialysis. Data shows that compared to angiotensin II more Hp is retained in the dialysis bag suggesting that there is increased aggregation of Hp. Data is the mean ± SE of three experiments