Recombinant Human UCH-L1/PGP9.5 Protein, CF

R&D Systems | Catalog # 6007-CY

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human UCH-L1/PGP9.5 Protein (6007-CY)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

P09936

Structure / Form

Monomer

Applications

Enzyme Activity
View all UCH-L1/PGP9.5 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human UCH-L1/PGP9.5 protein
Gln2-Ala223 with an N-terminal Met and 6-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

26 kDa

SDS-PAGE

27 kDa, reducing conditions

Activity

Measured by the hydrolysis of Ubiquitin-AMC.
The specific activity is >100 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6007-CY
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, Glycerol and DTT.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: UCH-L1/PGP9.5

Deubiquitination is a critical regulatory process in the ubiquitin-proteasome pathway (1). Ubiquitin C-terminal hydrolases (UCHs) are a family of cysteine proteases that catalyze the hydrolysis of a peptide bond at the C-terminal glycine of ubiquitin. Members of the UCH family have been implicated in a number of human diseases, including neurodegenerative diseases and cancers (2). Mutations of the UCH-L1 gene and alterations of the protein activity have been found to be associated with several neurodegenerative disorders, including Parkinson’s, Huntington’s and Alzheimer’s diseases (3).  It is also implicated in cancer tumorigenesis, including lung, breast, liver, kidney, colorectal and ovarian cancers (4-8). UCH-L1 is thought to be a tumor suppressor and biomarker for hepatocellular carcinoma and other digestive tumors.

References

  1. Wing, S. (2003) Int. J. Biochem. Cell Biol. 35:590.
  2. Ventii, KH and Wilkinson, KD (2008) Biochem. J. 414:161.
  3. Gong, B. and Leznik, E. (2007) Drug News Perspect. 20:365.
  4. Kim, H. et al. (2009) Oncogene 28:117.
  5. Wang, W. et al. (2008) Int. J. Oncol. 33:1037.
  6. Yu, J. et al. (2008) Hepatology 48:508.
  7. Kagara, I. et al. (2008) J. Urol. 180:343.
  8. Okochi-Takada, E. et al. (2006) Int. J. Cancer 119:1338.

Long Name

Ubiquitin C-terminal Hydrolase L1

Alternate Names

PARK5, PGP9.5, UCHL1

Entrez Gene IDs

7345 (Human); 22223 (Mouse)

Gene Symbol

UCHL1

UniProt

P09936 (Human)

Additional UCH-L1/PGP9.5 Products

Product Documents for Recombinant Human UCH-L1/PGP9.5 Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human UCH-L1/PGP9.5 Protein, CF

For research use only

Citations for Recombinant Human UCH-L1/PGP9.5 Protein, CF

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Protocols

View specific protocols for Recombinant Human UCH-L1/PGP9.5 Protein, CF (6007-CY):

Materials
  • Assay Buffer: 50 mM HEPES, 0.5 mM EDTA, 1 mM DTT, 0.1 mg/mL Ovalbumin, pH 8.0
  • Recombinant Human UCH-L1/PGP9.5 (rhUCH-L1) (Catalog # 6007-CY)
  • Substrate: Ubiquitin-AMC, (Boston Biochem, Catalog # U-550), 50 µM in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhUCH-L1 to 0.5 µg/mL in Assay Buffer.
  2. Dilute Substrate  to 2 µM in Assay Buffer.
  3. Load into a plate 50 µL of 0.5 µg/mL rhUCH-L1. Separately, also load 50 µL Assay Buffer to be used as a Substrate Blank.
  4. Seal plate and incubate both it and the diluted Substrate at 37 °C for 10 minutes. Also warm the plate reader to 37 °C.
  5. Start the reaction by adding 50 µL of 2 µM Substrate to all wells.
  6. Read at excitation and emission wavelengths of 380 nm and 460 nm, respectively, in kinetic mode for 5 minutes.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard 7-Amino, 4-Methyl Coumarin (Sigma, Catalog # A9891).

Per Well:

  • rhUCH-L1: 0.025 µg
  • Substrate: 1 µM

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