Procedure 2 Renaturing Fresh Phycocyanin Subunits

1. Dilute PC subunits from procedure 1 to 0.1 to 0.4 mg/mL protein with 8 M

Figure 2 Absorption spectra of the individual renatured a and P sub-units of phycocyanin purified from Synechococcus sp. PCC 6301.

Spectra were determined in 50 mM Na phosphate, 1 mM P-mercap-toethanol, pH 7.0, and at a protein concentration of 1.05 x 10-5 M for the a-subunit and 1.11 x 10-5 M for the P subunit. The Xmax was 620 nm for the a subunit and 608 nm for the P subunit. This figure was modified with permission from Reference 32.

Figure 2 Absorption spectra of the individual renatured a and P sub-units of phycocyanin purified from Synechococcus sp. PCC 6301.

Spectra were determined in 50 mM Na phosphate, 1 mM P-mercap-toethanol, pH 7.0, and at a protein concentration of 1.05 x 10-5 M for the a-subunit and 1.11 x 10-5 M for the P subunit. The Xmax was 620 nm for the a subunit and 608 nm for the P subunit. This figure was modified with permission from Reference 32.

Table 1. Millimolar Extinction Coefficients of urea, 5 mM P-mercaptoethanol, pH 8.0.

2. Dialyzed against 3 M urea, 5 mM P-mercaptoethanol, 6 mM Na-phos-phate, pH 6.7, at 4°C.

3. Dialyze against two changes of 10 mM Na-phosphate, 5 mM P-mercapto-ethanol, pH 6.5, at 4°C.

4. Dialyze against 5 mM Na-phosphate, pH 7.0, at 4°C.

0 0

Post a comment