Recombinant Human Kallikrein 14 Protein, CF

Name: Recombinant Human Kallikrein 14 Protein, CF
Brand: R&D Systems
SKU: 2626-SE
Rating: 5 (1 reviews)

Catalog #: 2626-SE
6 Citations 1 Review Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
2626-SE-010

Recombinant Human Kallikrein 14 Protein Enzyme Activity

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All Kallikrein 14 Proteins and Enzymes

Product Details

Citations (6)

FAQs

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Reviews (1)

Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Kallikrein 14 Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

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

Activity

Measured by its ability to cleave the fluorogenic peptide substrate Boc-VPR-AMC (Catalog # ES011). The specific activity is >3,000 pmol/min/µg, as measured under the described conditions.

Source

Mouse myeloma cell line, NS0-derived human Kallikrein 14 protein
Gln19-Met248, with a C-terminal 10-His tag

Accession #

AAD50773

N-terminal Sequence
Analysis

No results obtained: Gln19 predicted

Structure / Form

Pro form

Predicted Molecular Mass

26 kDa

SDS-PAGE

28 kDa, reducing conditions

Product Datasheets

2626-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.

2626-SE

Formulation	Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
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, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
Assay Buffer: 50 mM Tris, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 8.0
Recombinant Human Kallikrein 14 (rhKLK14) (Catalog # 2626-SE)
Bacterial Thermolysin (Thermolysin) (Catalog # 3097-ZN)
EDTA (Sigma, Catalog # E-4884)
Substrate: BOC-Val-Pro-Arg-AMC (Catalog # ES011), 100 mM stock in DMSO
F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
Fluorescent Plate reader (Model: SpectramMax Gemini EM by Molecular Devices) or equivalent

Dilute rhKLK14 to 200 µg/mL in Activation Buffer.
Dilute Thermolysin to 20 µg/mL in Activation Buffer.
Combine 25 µL of diluted rhKLK14 and 25 µL of diluted Thermolysin.
Incubate at 37 °C for 1 hour.
Stop reaction with 50 µL of 100 mM EDTA.
Dilute incubated rhKLK14 mixture to 0.2 ng/µL in Assay Buffer.
Dilute Substrate to 200 µM in Assay Buffer.
Load 50 µL of the 0.2 ng/µL rhKLK14 mixture in a plate, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.
Read at excitation and emission 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 V_max* (RFU/min) x Conversion Factor** (pmol/RFU)
	amount of enzyme (µg)

*Adjusted for Substrate Blank

**Derived using calibration standard 7-Amino, 4-Methyl Coumarin (AMC) (Sigma, Catalog # A-9891).

Per Well:

rhKLK14: 0.01 µg
Substrate: 100 µM

Scientific Data

Enzyme Activity

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Recombinant Human Kallikrein 14 (Catalog # 2626-SE) is measured by its ability to cleave the fluorogenic peptide substrate Boc-VPR-AMC (Catalog # ES011).

Reconstitution Calculator

Background: Kallikrein 14

Human tissue kallikreins refer to a group of secreted serine proteases that are encoded by homologous genes clustering on chromosome 19q13.3-4. As a member of this family, human tissue Kallikrein 14 (hKLK14) is present in many tissues, with high levels in breast, skin, prostate, and brain. The 251 amino acid hKLK14 precursor consists of a signal peptide (residues 1 to 18), a pro peptide (residues 19 to 24) and an active protein (residues 25 to 251) (1). Its enzymatic activity has been shown to be mainly trypsin-like (2). However, its physiological substrates and functions are still unclear. Several studies have suggested that hKLK14 may have clinical utility as a biomarker for cancer of the breast, ovary, and prostate (3, 4). In addition, it may be the initiator of a kallikrein proteolytic cascade responsible for the degradation of the adhesion structures in the stratum corneum (2). The purified, secreted rhKLK14 corresponds to the pro form (residues 19 to 248) with a replacement of the last three residues with a His tag at the C-terminus. When activated by thermolysin, it displays enzymatic activity towards a fluorogenic peptide described above. This activity can be inhibited by recombinant human Serpin A4, E1, and F2 (Catalog # 1669-PI, 1786-PI, and 1470-PI, respectively).

References

Yousef, G. M. et al. (2001) Cancer Res. 61:3425.
Brattsand, M. et al. (2004) J. Invest. Dermatol. 124:198.
Borgono, C. et al. (2003) Cancer Res. 63:9032.
Yousef, G. M. et al. (2003) Prostate. 56:287.

Entrez Gene IDs

43847 (Human)

Alternate Names

EC 3.4.21; EC 3.4.21.-; hK14; Kallikrein 14; Kallikrein-like protein 6; kallikrein-related peptidase 14; KLK14; KLKL6; KLK-L6kallikrein-14

Related Research Areas

Citations for Recombinant Human Kallikrein 14 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.

6 Citations: Showing 1 - 6
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Cross-linking of SPINK6 by transglutaminases protects from epidermal proteases.
Authors: Fischer J, Koblyakova Y, Latendorf T, Wu Z, Meyer-Hoffert U
J Invest Dermatol, 2013-01-10;133(5):1170-7.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay
Proteolytic activation cascade of the Netherton syndrome-defective protein, LEKTI, in the epidermis: implications for skin homeostasis.
Authors: Fortugno P, Bresciani A, Paolini C, Pazzagli C, El Hachem M, D'Alessio M, Zambruno G
J Invest Dermatol, 2011-06-23;131(11):2223-32.
Species: Human
Sample Types: Substrate
Applications: Enzyme Assay
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
Isolation of SPINK6 in human skin: selective inhibitor of kallikrein-related peptidases.
Authors: Meyer-Hoffert U, Wu Z, Kantyka T, Fischer J, Latendorf T, Hansmann B, Bartels J, He Y, Glaser R, Schroder JM
J. Biol. Chem., 2010-07-28;285(42):32174-81.
Species: Human
Sample Types: Peptide
Applications: Enzyme Assay
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
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