VYLSECKTGN GKNYRGTMSK TKNGITCQKW SSTSPHRPRF SPATHPSEGL EENYCRNPDN DPQGPWCYTT DPEKRYDYCD ILECEEECMH CSGENYDGKI SKTMSGLECQ AWDSQSPHAH GYIPSKFPNK NLKKNYCRNP DRELRPWCFT TDPNKRWELC DIPRCTTPPP SSGPTYQCLKGTGENYRGNV AVTVSGHTCQ HWSAQTPHTH NRTPENFPCK NLDENYCRNP DGKRAPWCHT TNSQVRWEYC KIPSCDSSP.
The activity is assayed on anti-proliferation and anti-migration of endothelial cells in vitro and anti-angiogenesis in vivo. The specific activity of anti-migration of endothelial cells in vitro is 55,000 Units/mg.
Lyophilized from a 0.2um filtered concentrated (1.0mg/ml) solution in 20mM NaAc, pH5.5, 4% mannitol.
Sterile Filtered White lyophilized (freeze-dried) powder.
Protein quantitation was carried out by two independent methods:1. UV spectroscopy at 280 nm using the absorbency value of 2.08 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).
2. Analysis by RP-HPLC, using a standard solution of Ang K1-3 as a Reference Standard.
Greater than 95.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Analysis by SDS-PAGE.
We recommend to briefly centrifuge the vial prior to opening to bring the contents to the bottom. Reconstitute in sterile distilled water or aqueous buffer containing 0.1% BSA to a concentration of 0.1-1.0 mg/ml. Stock solutions should be apportioned into working aliquots and stored at <-20oC. Further dilutions should be made in appropriate buffered solutions.
The lyophilized Angiostatin K1-3 is stable for several weeks at 2-8oC, but should be kept at -20oC for long term storage, preferably desiccated. Upon reconstitution, the preparation is stable for up to one week at 2-8oC. For maximal stability, apportion the reconstituted preparation into working aliquots and store at -20oC to -70oC.
Avoid repeated freeze/thaw cycles.
Angiostatin, Angiostatin Kringles 1-3, Ang K1-3.