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Recombinant Human Kallikrein 11 Protein, CF

Catalog #: 1595-SE
4 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
1595-SE-010
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Citations (4)
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Recombinant Human Kallikrein 11 Protein, CF Summary

Product Specifications

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.
Activity
Measured by its ability to cleave a colorimetric peptide substrate, N-carbobenzyloxy-Lys-ThioBenzyl ester (Z-Lys-SBzl), in the presence of 5,5’Dithio-bis (2-nitrobenzoic acid) (DTNB). Edwards, K.M. et al. (1999) J. Biol. Chem. 274:30468. The specific activity is >200 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Kallikrein 11 protein
Glu19-Asn250, with an N-terminal signal peptide and a C-terminal 10-His tag
Accession #
Q0WXX5
N-terminal Sequence
Analysis
Glu19
Structure / Form
Pro form
Predicted Molecular Mass
27 kDa
SDS-PAGE
40 kDa, reducing conditions

Product Datasheets

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1595-SE

Product Datasheet
COA
SDS

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

1595-SE

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and CaCl2.
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 opening.

Assay Procedure

Materials
  • Activation Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl, pH 7.5 (TCN)
  • Assay Buffer: 50 mM Tris, 1 M NaCl, 10 mM EDTA, pH 8.5
  • Recombinant Human Kallikrein 11 (rhKLK11) (Catalog # 1595-SE)
  • Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
  • EDTA (Sigma, Catalog # E-4884)
  • Fluorogenic Peptide Substrate: Z-Lys-SBzl (Bachem, Catalog # M-1300), 10 mM stock in DMSO
  • 5,5’-dithio-bis (2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D-8130), 10 mM stock in DMSO
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhKLK11 to 200 µg/mL in Activation Buffer.
  2. Dilute Thermolysin to 20 μg/mL in Activation Buffer.
  3. Combine equal volumes of 200 µg/mL rhKLK11 and 20 µg/mL Thermolysin.
  4. Incubate at 37 °C for 15 minutes.
  5. Stop Thermolysin activity by adding EDTA to a final concentration of 10 mM.
  6. Dilute incubated rhKLK11 to 2 ng/µL in Assay Buffer.
  7. Prepare Substrate Mixture containing 600 μM Substrate and 200 μM DTNB in Assay Buffer.
  8. Load into a clear plate 50 µL of the 2 ng/µL rhKLK11, and start the reaction by adding 50 µL of Substrate Mixture. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of Substrate Mixture.
  9. Read at a wavelength of 405 nm (bottom read) in kinetic mode for 5 minutes.
  10. 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 13260 M-1cm-1 
     ***Using the path correction 0.320 cm
     Note: the output of many spectrophotometers is in mOD Per Well:
  • rhKLK11: 0.1 µg
  • Substrate: 300 µM
  • DTNB: 100 µM
Reconstitution Calculator

Reconstitution Calculator

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 11

As a member of human tissue kallikrein family, Kallikrein 11, also known as hippostasin, trypsin-like serine protease and PRSS20, is encoded by the KLK11 gene (1). Two alternatively spliced forms exist, resulting in 250 (isoform 1) and 282 (isoform 2) amino acid sequences,respectively (2-5). Isoform 1 consists of a signal peptide (residues 1-18), a short pro peptide (residues 19-21) and the mature chain (residues 22-250). Isoform 2 is identical to isoform 1, except that a 32 amino acid segment is inserted in isoform 2 before residue 1 in isoform 1. Isoform 1 is predominantly expressed in brain whereas isoform 2 is preferentially expressed in prostate. KLK11 is a novel marker for ovarian and prostate cancer carcinomas (6-8). Recombinant human KLK11, after being activated by thermolysin, is active against a thioester substrate. This activity can be inhibited by AEBSF (Catalog # EI001), dichloroisocoumarin, and aprotinin. Recombinant human KLK11 produced by R&D Systems corresponds to isoform 1.

References
  1. Yousef, G.M. and E.P. Diamandis (2001) Endocrine Rev. 22:184.
  2. Yoshida, S. et al. (1998) Biochim. Biophys. Acta 1399:225. 
  3. Yousef, G.M. et al. (2000) Genomics 63:88.
  4. Mitsui, S. et al. (2000) Biochem. Biophys. Res. Commun. 272:205.
  5. Gan, L. et al. (2000) Gene 257:119.
  6. Diamandis, E.P. et al. (2002) Cancer Res. 62:295.
  7. Nakamura, T. et al. (2003) Urology 61:1042.
  8. Borgono, C.A. et al. (2003) Int. J. Cancer 106:605.
Entrez Gene IDs
11012 (Human)
Alternate Names
EC 3.4.21; EC 3.4.21.-; EC 3.4.21.4; Hippostasin; hK11; Kallikrein 11; kallikrein-related peptidase 11; KLK11; MGC33060; PRSS20kallikrein-11; Serine protease 20; TLSPhippostasin; Trypsin-like protease

Citations for Recombinant Human Kallikrein 11 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.

4 Citations: Showing 1 - 4
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  1. Inhibition of kallikrein-related peptidases by the serine protease inhibitor of Kazal-type 6.
    Authors: Kantyka T, Fischer J, Wu Z, Declercq W, Reiss K, Schroder J, Meyer-Hoffert U
    Peptides, 2011-03-23;32(6):1187-92.
    Applications: Bioassay
  2. Specific inhibition of tissue kallikrein 1 with a human monoclonal antibody reveals a potential role in airway diseases.
    Authors: Sexton DJ, Chen T, Martik D, Kuzmic P, Kuang G, Chen J, Nixon AE, Zuraw BL, Forteza RM, Abraham WM, Wood CR
    Biochem. J., 2009-08-13;422(2):383-92.
    Species: Human
    Sample Types: Antibody
    Applications: Surface Plasmon Resonance
  3. Activation profiles and regulatory cascades of the human kallikrein-related peptidases.
    Authors: Yoon H, Laxmikanthan G, Lee J, Blaber SI, Rodriguez A, Kogot JM, Scarisbrick IA, Blaber M
    J. Biol. Chem., 2007-09-06;282(44):31852-64.
    Applications: Enzyme Assay
  4. Enzymatic properties of human kallikrein-related peptidase 12 (KLK12).
    Authors: Memari</LastName><ForeNam N</Initial, Memari N, Jiang W, Diamandis EP, Luo LY
    Biol. Chem., 2007-04-01;388(4):427-35.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Enzyme Assay

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