| Species : |
Human |
| Source : |
E.coli |
| Tag : |
Non |
| Protein Length : |
1-321 a.a. |
| Description : |
The protein coding region of the catalytic domain of PTPN1 (amino acids 1-321) was cloned into an E. coli expression vector. The catalytic domain of PTP-1B was overexpressed in E. coli as a soluble protein. |
| Form : |
Liquid. In 25 mM Tris-HCl buffer (pH 7.5) containing 1 mM DTT, 2 mM beta-mercaptoethanol, 1 mM EDTA, 20%Glycerol |
| Molecular Weight : |
37.3 kDa (321 aa) |
| Purity : |
> 95% by SDS - PAGE |
| Concentration : |
1 mg/ml (determined by Bradford assay) |
| Biological activity : |
Specific activity: 6,000-12,000units/mg. Enzymatic activity was confirmed by measuring the amount of enzyme hydrolyzing 1 nmole of p-nitrophenyl phosphate (pNPP) per minute at 37°C, pH7.5 using 10mM of substrate. |
| Activity Assay : |
1. Prepare a 200ul reaction mix into a suitable container: The final concentrations are 10mM Tris(pH7.5), 50mM NaCl, 2mM DTT, 1mM MnCl2, 10mM pNPP. 2. Pre-warm reaction mix in heat block at 37°C 3. Add recombinant PTP1B solution with various concentrations (2ug, 3ug, 4ug) in 200ul reaction buffer. 4. Mix by inversion and Incubate at 37°C for 10minutes. 5. Stop the reaction by adding the 800ul of 0.1N NaOH 6. Record the absorbance at 405nm |
| Sequences of amino acids : |
MEMEKEFEQI DKSGSWAAIY QDIRHEASDF PCRVAKLPKN KNRNRYRDVS PFDHSRIKLH QEDNDYINAS LIKMEEAQRS YILTQGPLPN TCGHFWEMVW EQKSRGVVML NRVMEKGSLK CAQYWPQKEE KEMIFEDTNL KLTLISEDIK SYYTVRQLEL ENLTTQETRE ILHFHYTTWP DFGVPESPAS FLNFLFKVRE SGSLSPEHGP VVVHCSAGIG RSGTFCLADT CLLLMDKRKD PSSVDIKKVL LEMRKFRMGL IQTADQLRFS YLAVIEGAKF IMGDSSVQDQ WKELSHEDLE PPPEHIPPPP RPPKRILEPH N |
| 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 |