| Cat. No. : |
PIN1-351H |
| Description : |
Human Pin 1 is a peptidyl-prolyl cis/trans isomerase (PPIase) that interacts with NIMA and essential for cell cycle regulation Pin1 is nuclear PPIase containing a WW protein interaction domain, and is structurally and functionally related to Ess1/Ptf1, an essential protein in budding yeast. PPIase activity is necessary for Ess1/Pin1 function in yeast. Pin1 is thus an essential PPIase that regulates mitosis presumably by interacting with NIMA and attenuating its mitosis-promoting activity. Substrates of Pin1 include the mitotic regulators (Cdc25 phosphatase and NIMA ,PLK I, Wee, and Myt1 kinases), several transcription factors like Catenin, c-Jun, and the tumor suppressor protein p53 , and some specific proteins like the RNA Pol II, the cytoskeleton protein tau, and the G1/S protein Cyclin D1. |
| Source : |
Escherichia Coli. |
| Form : |
Liquid. In 20 mM Tris-HCl buffer (pH 7.5) containing 100 mM NaCl, 5 mM DTT, 20% glycerol. |
| Purity : |
> 95% by SDS - PAGE. |
| Concentration : |
1 mg/ml (determined by Bradford assay) |
| Endotoxin Level : |
< 1.0 EU per 1 μg of protein (determined by LAL method) |
| Sequences of amino acids : |
MADEEKLPPG WEKRMSRSSG RVYYFNHITN ASQWERPSGN SSSGGKNG QG E PARVRCSHL LVKHSQSRRP SSWRQEKITR TKEEALELINGYIQKIKSGE EDFESLASQF SDCSSAKARG DLGAFSRGQM QKPFEDASFA LRTGEMSG PV FTDSGIHIIL RTE |
| Biological activity : |
Specific activity is > 330 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 PIN 1 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 +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. |
