KLK5/Kallikrein 5 His Tag Protein, Human
KLK5/Kallikrein 5 His Tag Protein, Human
Product Details
Product Details
Product Specification
| Species | Human |
| Synonyms | Kallikrein-5, SCTE, Kallikrein-like protein 2 |
| Accession | NP_036559.1 |
| Amino Acid Sequence | Val23-Ser293 with His Tag at the C-Terminus |
| Expression System | HEK293 |
| Molecular Weight | 35-40kDa (Reducing) |
| Purity | ＞95% by SDS-PAGE |
| Conjugation | Unconjugated |
| Tag | His Tag |
| Physical Appearance | Lyophilized Powder |
| Storage Buffer | 50 mM MES, 150 mM NaCl, pH 5.5 |
| Reconstitution | Reconstitute at 0.1-1 mg/ml according to the size in ultrapure water after rapid centrifugation. |
| Stability & Storage |
· 12 months from date of receipt, lyophilized powder stored at -20 to -80℃. · 3 months, -20 to -80℃ under sterile conditions after reconstitution. · 1 week, 2 to 8℃ under sterile conditions after reconstitution. · Please avoid repeated freeze-thaw cycles. |
| Reference | 1.Palacios Serrato EG, Medina-Abreu KH, Oropeza-Martínez E, Jacinto-Alemán LF, Macías-Silva M, Tecalco-Cruz AC. ADAMTS-8 and kallikrein-related peptidases 10 and 5 proteases also have a tumor suppression role. World J Clin Oncol. 2025 Nov 24;16(11):110202. |
Background
KLK5 protein (kallikrein-related peptidase 5) is a secreted serine protease encoded by the KLK5 gene, featuring a signal peptide and a catalytically active trypsin-like domain in its structure, with predominant expression in epithelial tissues such as skin and salivary glands. Physiologically, KLK5 participates in epidermal desquamation by cleaving cell adhesion molecules and activates antimicrobial peptide precursors to maintain skin barrier homeostasis, while also serving as a key host protease that cleaves coronavirus spike proteins to facilitate viral entry. Clinically, aberrant expression of KLK5 is closely associated with multiple diseases: its dysregulated activity can lead to inflammatory skin disorders including Netherton syndrome, atopic dermatitis, and rosacea; in oncology, it exhibits a complex dual role—with high expression in ovarian cancer indicating poor prognosis, whereas low expression in testicular cancer correlates with advanced disease progression. In recent years, it has also been linked to low type 2 inflammation-associated asthma, establishing KLK5 as an important therapeutic target for the development of anti-coronavirus and anti-inflammatory skin disease drugs.
Protocol
Assay protocol
Principle: Measured by its ability to cleave the fluorogenic peptide substrate, Boc-VPR-AMC.
Materials
1.Assay Buffer: 0.1 M NaH2PO4, pH 8.0
2.KLK5/Kallikrein 5 His Tag Protein, Human
3.Substrate: Fluorogenic Peptide Substrate BOC-Val-Pro-Arg-AMC (R&D systems, Catalog # ES011)
4.96 ELISA Removable Plate, Black, High binding (GENEVER, Catalog # GMO2-96H)
5.Plate Reader (PerkinElmer, excitation 380 nm and emission 460 nm)
Produce
1.Dilute Kallikrein 5 to 8, 4 and 2 μg/mL in Assay Buffer.
2.Dilute Substrate to 200 μM with Assay Buffer.
3.Load 50 μL of the dilute Kallikrein 5 in a plate, and start the reaction by adding 50 μL of 200 μM Substrate. Include a Substrate Blank containing 50 μL assay buffer and 50 μL of 200 μM Substrate.
4.Read in kinetic mode for 10 minutes at excitation and emission wavelengths of 380 nm and 460 nm.
Standard Curve
1.Dilute Calibration standard to 10 μM in Assay Buffer and prepare serial dilutions.
2.Add 100 μL of each serially diluted standard and blank (Assay buffer) into appropriate wells of a 96-well, the standard curve has a range of 1000, 500, 250, 125, 62.5, 31.3, 15.6, 7.8 pmol per well.
3.Read at excitation and emission wavelengths of 380 nm and 460 nm, respectively in endpoint mode.
4.Linear Regression of 7-amino-4-Methyl Coumarin (pmol)(y) –RFU-Blank(x):
Specific Activity (pmol/min/µg) = |
Adjusted Vmax* (RFU/min) x Conversion Factor (pmol/RFU) |
amount of enzyme (µg) |
*Adjusted for Substrate Blank
