12-Lipoxygenase (porcine leukocyte)

Molecular Formula : Monomer
Stability : 1 year
Formulation : A solution in 20 mM phosphate buffer, pH 7.5
Description :
12-Lipoxygenase was purified from porcine leukocytes as described (12). Purified enzyme (0.25 mg) was digested with Achromobacterproteinase I (3 ,g) in 170 ,ul of 0.1 M Tris HCl(pH 8.0) containing 0.1% NaDodSO4 at 37°C for 24 hr. After
digestion, the peptides were purified using a Hitachi HPLC system with a reverse-phase column (SynChropak, 4 x 250mm;Alltech) at a flow rate of 1 ml/min. Material was eluted with a 25-50o (vol/vol) acetonitrile gradient in 0.1% trifluoroacetic acid over 50 min, and the absorbance at 210 nm of the eluate was continuously monitored. Amino-terminal sequences of the native enzyme and its peptide fragments were analyzed using a gas-phase sequencer model 470A and a phenylthiohydantoin amino acid derivative analyzer model 120A (Applied Biosystems). For the determination of amino acid composition, the purified enzyme (8 jig) was hydrolyzed in vacuo at 110°C for 20 hr with 6 M HCI, and amino acid analysis was carried out using a Waters PicoTag system.
Purity : cell lysate 100,000 x g supernatant
Storage : -80°C