Recombinant Human Tripeptidyl-peptidase I/TPP1 Protein, CF

R&D Systems | Catalog # 2237-SE

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems NS0-derived Recombinant Human Tripeptidyl-peptidase I/TPP1 Protein (2237-SE)
  • 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

NS0

Accession Number

O14773

Applications

Enzyme Activity
View all Tripeptidyl-Peptidase I/TPP1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Tripeptidyl-Peptidase I/TPP1 protein
Met1-Pro563, with a C-terminal 10-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

N-terminal Sequence Analysis

Ser20

Predicted Molecular Mass

61 kDa

SDS-PAGE

65-75 kDa, reducing conditions

Activity

Measured by the cleavage of Ala-Ala-Phe-AMC.
The specific activity is >850 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

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).

References

  1. Sleat, D.E. et al. (1997) Science 277:1802.
  2. Ezaki, J. et al. (1999) J. Neurochem. 72:2573.
  3. Tomkinson, B. (1999) Trends Biochem. Sci. 24:355.
  4. Lin, L. et al. (2001) J. Biol. Chem. 276:2249.
  5. Kida, E. et al. (2001) J. Neuropathol. Exp. Neurol. 60:280.
  6. Sleat, D.E. et al. (1999) Am. J. Hum. Genet. 64:1511.         

Alternate Names

CLN2, LINCL, LPIC, TPP1, TripeptidylPeptidase I

Entrez Gene IDs

1200 (Human)

Gene Symbol

TPP1

UniProt

O14773

Additional Tripeptidyl-Peptidase I/TPP1 Products

Product Documents for Recombinant Human Tripeptidyl-peptidase I/TPP1 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.

Download SDS

Product Specific Notices for Recombinant Human Tripeptidyl-peptidase I/TPP1 Protein, CF

For research use only

Related Research Areas

Serine Proteases and Regulators

Citations for Recombinant Human Tripeptidyl-peptidase I/TPP1 Protein, CF

Customer Reviews for Recombinant Human Tripeptidyl-peptidase I/TPP1 Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human Tripeptidyl-peptidase I/TPP1 Protein, CF and earn rewards!

Have you used Recombinant Human Tripeptidyl-peptidase I/TPP1 Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant Human Tripeptidyl-peptidase I/TPP1 Protein, CF (2237-SE):

Materials
  • 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
  1. Dilute rhTPP1 to 20 μg/mL in Activation Buffer.
  2. Incubate for 1 hour at 37 °C.
  3. Dilute rhTPP1 to 0.5 ng/μL in Assay Buffer.
  4. Dilute Substrate to 400 µM in Assay Buffer.
  5. 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.
  6. Read at excitation and emssion wavelengths of 380 nm and 460 nM (top read), 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 # A-9891).

Per Well:
  • rhTPP1: 0.025 µg
  • Substrate: 200 µM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes