Recombinant Human Kallikrein 3/PSA Protein, CF

R&D Systems | Catalog # 1344-SE

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Kallikrein 3/PSA Protein (1344-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

P07288

Structure / Form

Pro form

Applications

Enzyme Activity
View all Kallikrein 3/PSA Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Kallikrein 3/PSA protein
Ala18-Pro261, 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

Ala18

Predicted Molecular Mass

28 kDa

SDS-PAGE

36 kDa, reducing conditions

Activity

Measured by its ability to cleave the colorimetric peptide substrate, Succinyl-Arg-Pro-Tyr-p-Nitroanilide (Suc-RPY-pNA).
The specific activity is >70 pmol/min/µg, as measured under the described conditions.

Reviewed Applications

Read 2 reviews rated 5 using 1344-SE in the following applications:

Scientific Data Images for Recombinant Human Kallikrein 3/PSA Protein, CF

Recombinant Human Kallikrein 3/PSA Protein Enzyme Activity

Recombinant Human Kallikrein 3/PSA Protein Enzyme Activity

Recombinant Human Kallikrein 3 (Catalog # 1344-SE) is measured by its ability to cleave the colorimetric peptide substrate, Succinyl-Arg-Pro-Tyr-p-Nitroanilide (Suc-RPY-pNA).

Formulation, Preparation, and Storage

1344-SE
Formulation Lyophilized from a 0.2 μm filtered solution in Tris, NaCl and CaCl2.
Reconstitution Reconstitute at 200 μg/mL in 50 mM Tris, 10 mM CaCl2,150 mM NaCl and 0.05% (w/v) Brij-35, pH 7.5.
Shipping The product is shipped with polar packs. 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, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Kallikrein 3/PSA

Kallikrein 3, commonly known as prostate specific antigen (PSA), is a serine protease of the human tissue Kallikrein gene family (1). PSA is synthesized in the ductal and acinar epithelium of the prostate gland and secreted into the seminal plasma in high concentrations (0.5 - 2 g/L) (2). A small portion of PSA “leaks” into the systemic circulation, the levels of which increase significantly (30-fold) from prostate cancer tissue than normal prostate tissue (3). PSA has become a well established tumor marker that aids the diagnosis, staging, and follow up of prostate cancer.

The deduced amino acid sequence of human PSA consists of a signal peptide, a short pro region and a mature/active enzyme. The pro-enzyme is activated, possibly by active Kallikreins 2, 4 or 15 in vivo (4). rhPSA is activated by thermolysin, a zinc protease. The active PSA cleaves several tyrosyl peptide bonds in semenogelins I and II, which are the major gel-forming proteins produced by the seminal vesicles (5). Several inhibitors including serpin A3/ alpha 1-antichymotrypsin (ACT) and alpha 2‑macroglobulin are known to form complexes with PSA.

References

  1. Yousef, G.M. and E.P. Diamandis (2001) Endocrine Rev. 22:184.
  2. Ward, A.M. et al. (2001) Ann. Clin. Biochem. 38:633.
  3. Jain, S. et al. (2002) Postgrad. Med. J. 78:646.
  4. LiLja H. (2003) Urology 62:27.
  5. Takayama, T.K. et al. (1997) J. Biol. Chem. 272:21582.

Alternate Names

KLK3, PSA

Entrez Gene IDs

354 (Human)

Gene Symbol

KLK3

UniProt

P07288

Additional Kallikrein 3/PSA Products

Product Documents for Recombinant Human Kallikrein 3/PSA Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Kallikrein 3/PSA Protein, CF

For research use only

Citations for Recombinant Human Kallikrein 3/PSA Protein, CF

Customer Reviews for Recombinant Human Kallikrein 3/PSA Protein, CF (2)

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Protocols

View specific protocols for Recombinant Human Kallikrein 3/PSA Protein, CF (1344-SE):

Materials
  • Activation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, 0.05% Brij-35, pH 7.5 (TCNB)
  • Assay Buffer: 50 mM Tris, 1 M NaCl, pH 8.0
  • Recombinant Human Kallikrein 3/PSA (rhKLK3) (Catalog # 1344-SE)
  • Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
  • 1,10 Phenanthroline (Sigma, Catalog # 320056), 0.6 M stock in DMSO
  • Substrate: Suc-Arg-Pro-Tyr-pNa (AnaSpec, Catalog # 20586), 10 mM in deionized water
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhKLK3 to 200 µg/mL in Activation Buffer.
  2. Dilute Thermolysin to 2 µg/mL in Activation Buffer.
  3. Combine equal volumes of 200 µg/mL rhKLK3 and 2 µg/mL Thermolysin.
  4. Incubate at 37 °C for 5 minutes.
  5. Dilute 1,10 Phenanthroline to 20 mM in Assay Buffer.
  6. Stop Thermolysin activity  by adding 1,10 Phenanthroline to a final concentration of 10 mM.
  7. Dilute activated rhKLK3 to 20 ng/µL in Assay Buffer.
  8. Dilute Substrate to 2 mM in Assay Buffer.
  9. Load 50 µL of the 20 ng/µL rhKLK3 in a plate and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 2 mM Substrate.
  10. Read at a wavelength of 405 nm (bottom read) in kinetic mode for 5 minutes.
  11. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

     *Adjusted for Substrate Blank 
     **Using the extinction coefficient 8800 M-1cm-1 
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD. Per Well:

  • rhKLK3: 1 µg
  • Substrate: 1 mM

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