Recombinant Human Tripeptidyl-peptidase I/TPP1 Protein, CF
Recombinant Human Tripeptidyl-peptidase I/TPP1 Protein, CF Summary
Product Specifications
Met1-Pro563, with a C-terminal 10-His tag
Analysis
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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.
2237-SE
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| 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
- Activation Buffer: 50 mM NaOAc, 100 mM NaCl, pH 3.5
- Assay Buffer: 50 mM NaOAc, 100 mM NaCl, pH 5.0
- Recombinant Human Tripeptidyl‑Peptidase I/TPP1 (rhTPP1) (Catalog # 2237-SE)
- Substrate: AAF-AMC (Bachem, Catalog # I-1415), 10 mM in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: Gemini EM by Molecular Devices) or equivalent
- Dilute rhTPP1 to 20 μg/mL in Activation Buffer.
- Incubate for 1 hour at 37 °C.
- Dilute rhTPP1 to 0.5 ng/μL in Assay Buffer.
- Dilute Substrate to 400 µM in Assay Buffer.
- Load into a black well plate 50 µL of the 0.5 μg/mL rhTPP1 and start the reaction by adding 50 µL of 400 µM substrate.
- Read at excitation and emssion wavelengths of 380 nm and 460 nM (top read), 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 # A-9891).
- rhTPP1: 0.025 µg
- Substrate: 200 µM
Background: Tripeptidyl-Peptidase I/TPP1
Tripeptidyl-Peptidase I (TPP1), also known as tripeptidyl aminopeptidase, is a lysosomal peptidase which can hydrolyze tripeptides from the N-termini of oligopeptides and also possesses weak endopeptidase activity (1‑3). TPP1 is a serine peptidase with a Ser-Glu-Asp catalytic triad, making it a member of the sedolisin family (4). The TPP1 precursor undergoes autoactivation under conditions of acidic pH (4). TPP1 is expressed in many tissues, with elevated expression in tissues associated with peptide hormone production (5). Mutations in TPP1 have been shown to be a cause of classic late‑infantile neuronal ceroid lipofuscinosis (CLN2), a lysosomal storage disease (6).
- Sleat, D.E. et al. (1997) Science 277:1802.
- Ezaki, J. et al. (1999) J. Neurochem. 72:2573.
- Tomkinson, B. (1999) Trends Biochem. Sci. 24:355.
- Lin, L. et al. (2001) J. Biol. Chem. 276:2249.
- Kida, E. et al. (2001) J. Neuropathol. Exp. Neurol. 60:280.
- Sleat, D.E. et al. (1999) Am. J. Hum. Genet. 64:1511.
Citations for Recombinant Human Tripeptidyl-peptidase I/TPP1 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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Neural stem cells for disease modeling and evaluation of therapeutics for infantile (CLN1/PPT1) and late infantile (CLN2/TPP1) neuronal ceroid lipofuscinoses
Authors: N Sima, R Li, W Huang, M Xu, J Beers, J Zou, S Titus, EA Ottinger, JJ Marugan, X Xie, W Zheng
Orphanet J Rare Dis, 2018-04-10;13(1):54.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
Intravitreal implantation of TPP1-transduced stem cells delays retinal degeneration in canine CLN2 neuronal ceroid lipofuscinosis
Exp Eye Res, 2016-09-13;0(0):.
Species: Human
Sample Types:
Applications: Enzyme Assay
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