||Recombinant human cyclophilin H was expressed in E.coli and purified by conventional chromatography techniques, 19.2 kDa (177 aa).
||Cyclophilin H (also known as peptidylpropyl isomerase H, PPIH) is a member of peptidyl-propyl cis-trans isomerase (PPIase) family, which catalyzes the cis-transisomerization of proline imidic peptide bonds in oligopeptides and accelerates the folding of proteins. The cyclophilin H is a specific component of the human U4/U6 small nuclear ribonucleoprotein particle involved in the nuclear splicing of premRNA.It stably associates with the U4/U6-60kD and -90kD proteins, the human orthologues of the Saccharomyces cerevisiaePrp4 and Prp3 splicing factors.
|Sequences of amino acids:
||MAVANSSPVN PVVFFDVSIGG QEVGRMKIEL FADVVPKTAE NFRQFCTGEF RKDGVPIGYK GSTFHRVIKD FMIQGGDFVN GDGTGVASIY RGPFADENFK LRHSAPGLLS MANSGPSTN GCQFFITCSK CDWLDGKHV VFGKIIDGLL VMRKIENVPT GPNNKPKLPV V ISQCGEM
||Liquid in phosphate-buffered saline ( pH 7.4) containing 10% glycerol.
||> 95% by SDS PAGE
||1 mg/ml (determined by Bradford assay)
||Specific activity is > 220 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.
||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 Cyclophikin H (PPIH) 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.
||Can be stored at +4°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.