Cat. No. : |
PPIA-60H |
Description : |
PPIases catalyze the cis-trans isomerization of proline imidic peptide bonds in oligopeptides and accelerate the folding of proteins. Cyclophilin-A is a cyclosporin binding-protein and may play a role in cyclosporin A-mediated immunosuppression. Cyclophilin-A can also interact with several HIV proteins, including p55 gag, Vpr, and capsid protein, and has been shown to be necessary for the formation of infectious HIV virions. Multiple pseudogenes that map to different chromosomes have been reported. |
Source : |
Escherichia Coli. |
Amino Acid Sequence : |
MGSSHHHHHH SSGLVPRGSH MVNPTVFFDI AVDGEPLGRV SFELFADKVP KTAENFRALSTGEKGFGYKG SCFHRIIPGF MCQGGDFTRH NGTGGKSIYG EKFEDENFIL KHTGPGILSMANAGPNTNGS QFFICTAKTE WLDGKHVVFG KVKEGMNIVE AMERFGSRNG KTSKKITIADCGQLE. |
Physical Appearance : |
Sterile filtered colorless solution. |
Purity : |
Greater than 95.0% as determined by: (a) Analysis by RP-HPLC. (b) Analysis by SDS-PAGE. |
Formulation : |
1 mg/ml solution containing 20 mM Tris 8.0, 20 mM NaCl, 0.5 mM DTT and 10% glycerol. |
Stability : |
PPIase-A Human Recombinant although stable at 14°C for 1 week, should be stored desiccated below -18°C. Please prevent freeze thaw cycles. |
Biological activity : |
Specific activity is > 270 nmoles/min/mg, and is defined as the amount of enzyme that cleaves 1umole of suc-AAFP-pNA per minute at 25°C in Tris-Hcl pH8.0 using chymotrypsin. |
Activity Assay : |
1. Prepare 170ul assay buffer into a suitable container and pre-chill on ice before use: The final concentrations are 200 mM Tris-Hcl, pH 8.0, and 20nM chymotrypsin.2. Add 10ul of recombinant Cyclophilin A protein with 1ug in assay buffer. 3. Mix by inversion and equilibrate to 1°C and monitor the A405nm until the value is constant using a spectrophotometer.4. Add 20ul pre-chilled 5mM suc-AAFP-pNA. (Substrate was dissolved in TFE that contained 460mM LiCl to a concentration of 3 mM)5. Record the increase in A405 nm for 30 minutes at 25°C. |
Storage : |
Can be stored at +°C short term (1-2 weeks). For long term storage, aliquot and store at -20°C or -70°C. Avoid repeated freezing and thawing cycles. |