Recombinant Human Kallikrein 12 Protein, CF Summary
Product Specifications
Ala18-Asn248, with a C-terminal 10-His tag
Analysis
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.
3095-SE
| Formulation | Supplied as a 0.2 μm filtered solution in MES, 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.
|
Assay Procedure
- Activation Buffer: 100 mM Tris, 150 mM NaCl, 10 mM CaCl2, 0.05% (w/v) Brij-35, pH 8.0
- Assay Buffer: 100 mM Tris, 150 mM NaCl, 10 mM CaCl2, 0.05% (w/v) Brij-35, pH 7.5
- Recombinant Human Kallikrein 12 (rhKLK12) (Catalog # 3095-SE)
- Fluorescent Peptide Substrate: BOC-Val-Pro-Arg-AMC (Catalog # ES011), 10 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhKLK12 to 100 µg/mL in Activation Buffer.
- Incubate 100 µg/mL rhKLK12 at 37 °C for 24 hours.
- Dilute rhKLK12 to 0.2 ng/µL in Assay Buffer.
- Dilute Substrate to 200 µM in Assay Buffer.
- Load into plate 50 µL of 0.2 ng/µL rhKLK12, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing Assay Buffer in place of rhKLK12.
- Read plate 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 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 (AMC) (Sigma, Catalog # A-9891).
- rhKLK12: 0.01 µg
- Substrate: 100 µM
Reconstitution Calculator
Background: Kallikrein 12
Human tissue Kallikrein 12, encoded by the KLK12 gene, is a secreted serine protease that belongs to the human tissue kallikrein family. It is present in many tissues, such as salivary gland, stomach and breast. KLK12 displays trypsin-like enzymatic activity. This activity can be inhibited by Serpin F2 (Catalog # 1470-PI). The physiological functions of KLK12 still remain unclear. Its expression is modulated by steroid hormones and is down‑regulated in breast cancer (2). Human KLK12 has three splice variants resulting from alternative splicing of the 3’ end. The KLK12 produced by the R&D Systems corresponds to the full-length classical form, also known as isoform 2 (1). The amino acid sequence of human KLK12 is 80%, 77%, 71% and 66% to that of bovine, canine, mouse, and rat. The recombinant human KLK12 can be autoactivated under the conditions described in the Activity Assay Protocol. The active enzyme has the N-terminal sequence of I22FNGTECGRNS.
- Yousef, G.M. et al. (2000) Genomics. 69:331.
- Yousef, G.M. and E.P. Diamandis (2001) Endocrine Rev. 22:184.
Citations for Recombinant Human Kallikrein 12 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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A peptide-based approach to evaluate the adaptability of influenza A virus to humans based on its hemagglutinin proteolytic cleavage site
Authors: MR Straus, GR Whittaker
PLoS ONE, 2017-03-30;12(3):e0174827.
Species: Virus - Influenza A
Sample Types: Peptide
Applications: Enzyme Assay -
Angiogenesis stimulated by human kallikrein-related peptidase 12 acting via a platelet-derived growth factor B-dependent paracrine pathway.
Authors: Kryza T, Achard C, Parent C, Marchand-Adam S, Guillon-Munos A, Iochmann S, Korkmaz B, Respaud R, Courty Y, Heuze-Vourc'h N
FASEB J, 2013-11-13;28(2):740-51.
Applications: Enzyme Assay -
Kallikrein-related peptidase 12 hydrolyzes matricellular proteins of the CCN family and modifies interactions of CCN1 and CCN5 with growth factors.
Authors: Guillon-Munos A, Oikonomopoulou K, Michel N, Smith CR, Petit-Courty A, Canepa S, Reverdiau P, Heuzé-Vourc'h N, Diamandis EP, Courty Y
J. Biol. Chem., 2011-05-31;286(29):25505-18.
Species: Human
Sample Types: Recombinant Protein
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 -
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: Fluorogenic Peptide Substrate
Applications: Enzyme Assay
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