Source: The sequence (Accession # NP_001116849) corresponding to the full length human DJ-1 protein along with N-terminal GST tag was expressed in E. coli. Recombinant DJ-1 protein was purified from bacterial lysate using GST affinity chromatography. GST tag was not removed. The molecular weight of the recombinant GST_DJ-1 protein is 47.3 kDa.
Activity: GST DJ-1 was assayed for protease activity using SensoLyte® Green Protease Assay Kit (AnaSpec Cat# 71124).
Applications: ELISA, WB, Protease Activity Determination.
Purity: Greater than 95%, as determined by SDS-PAGE.
Storage: Purified GST-DJ-1 is supplied as sterile and frozen at 1 mg/ml in 50 mM Tris-Cl/150mM NaCl, pH=8.0. Store at 2-4ºC for immediate use within 1-2 weeks or at –80ºC for up to 12 months. Avoid repeated freeze-thaw cycles.
Description: DJ-1 (also known as PARK7, Parkinson disease protein 7) is a 189-amino acid protein that is ubiquitously expressed in many organs including brain, liver, kidney, pancreas, heart, and others.1-5 Although DJ-1 was originally discovered as a novel oncogene product, it was found to play several other roles in biological processes. These include regulation of RNA binding activity, fertility, anti-oxidative stress, and is linked to the early onset of Parkinson disease when mutated.1-5 Recently, it was demonstrated that DJ-1 can act as an atypical peroxiredoxin-like peroxidase5 and possesses protease activity.4
1. Huai, Q. et al. FEBS Letters 549, 171 (2003).
2. Inden, M. et al. J Pharma Sci 117, 189 (2011).
3. Kitamura, Y. et al. Mol Neurodegener 6, 1 (2011).
4. Chen, J. et al. Hum Mol Genetics 19, 2395 (2010).
5. Andres-Mateos, E., et al. Proc Nat Acad Sci 104, 14807 (2007).
Figure 1. Human GST-DJ-1 on SDS-PAGE. Purified GST-DJ-1 was loaded onto a 10-20% Tris-HCl gel at 3 μg/well and resolved at 200V for 40 min. 1=Uninduced cell lysate, 2=Induced cell lysate, 3=Purified GST- DJ-1 protein.
Figure 2. Western Immunoblot of DJ-1. Purified GST-DJ-1 protein was probed with anti-DJ-1 specific antibody.
Figure 3. GST-DJ-1 Proteolytic Activity. GST-DJ-1 was loaded at 200 µg/well with 1mM DTT and assayed for proteolytic activity using SensoLyte® Green Protease Assay Kit (AnaSpec Cat# 71124).