Recombinant Human UCH-L1/PGP9.5 Protein, CF Summary
Product Specifications
Gln2-Ala223 with an N-terminal Met and 6-His tag
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
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.
|
Assay Procedure
- 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
- Dilute rhUCH-L1 to 0.5 µg/mL in Assay Buffer.
- Dilute Substrate to 2 µM in Assay Buffer.
- 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.
- Seal plate and incubate both it and the diluted Substrate at 37 °C for 10 minutes. Also warm the plate reader to 37 °C.
- Start the reaction by adding 50 µL of 2 µM Substrate to all wells.
- Read at excitation and emission wavelengths of 380 nm and 460 nm, respectively, in kinetic mode for 5 minutes.
- 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).
- rhUCH-L1: 0.025 µg
- Substrate: 1 µM
Reconstitution Calculator
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.
- Wing, S. (2003) Int. J. Biochem. Cell Biol. 35:590.
- Ventii, KH and Wilkinson, KD (2008) Biochem. J. 414:161.
- Gong, B. and Leznik, E. (2007) Drug News Perspect. 20:365.
- Kim, H. et al. (2009) Oncogene 28:117.
- Wang, W. et al. (2008) Int. J. Oncol. 33:1037.
- Yu, J. et al. (2008) Hepatology 48:508.
- Kagara, I. et al. (2008) J. Urol. 180:343.
- Okochi-Takada, E. et al. (2006) Int. J. Cancer 119:1338.
Citations for Recombinant Human UCH-L1/PGP9.5 Protein, CF
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 2
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UCHL1, besides leptin and fibronectin, also could be a sensitive marker of the relapsing-remitting type of multiple sclerosis
Authors: E Górska, M Tylicka, A Hermanowic, E Matuszczak, A Sankiewicz, E Gorodkiewi, J Hermanowic, E Karpi?ska, K Socha, J Kochanowic, M Jakoniuk, J Kami?ska, E Homšak, OM Koper-Lenk
Scientific Reports, 2023-02-28;13(1):3423.
Species: Human
Sample Types:
Applications: SPR -
UCHL1 and Proteasome in Blood Serum in Relation to Dietary Habits, Concentration of Selected Antioxidant Minerals and Total Antioxidant Status among Patients with Alzheimer's Disease
Authors: S Bogdan, A Pu?cion-Ja, K Klimiuk, K Socha, J Kochanowic, E Gorodkiewi
Journal of Clinical Medicine, 2022-01-14;11(2):.
Species: Human
Sample Types: N/A
Applications: Surface Plasmon Resonance (SPR
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