Kallikrein-5 Protein, Human (HEK293, His, solution)

The Kallikrein-5 protein may be involved in desquamation, suggesting a role in the process of skin shedding and exfoliation.Its association with desquamation suggests that it plays a role in regulating the removal of dead skin cells, which is critical for skin homeostasis.Kallikrein-5 Protein, Human (HEK293, His, solution) is the recombinant human-derived Kallikrein-5 protein, expressed by HEK293 , with C-6*His labeled tag.

Kallikrein-5 Protein appears to play a potential role in desquamation, suggesting involvement in the intricate processes of skin shedding and exfoliation. Its potential connection to desquamation implies a functional role in regulating the removal of dead skin cells, a crucial aspect of skin homeostasis. Notably, Kallikrein-5 activity is inhibited by Zn2+, indicating a potential regulatory mechanism for its enzymatic function. Understanding the specific mechanisms through which Kallikrein-5 contributes to desquamation and the regulatory role of Zn2+ could provide valuable insights into its function in skin physiology and shed light on its potential significance in processes related to skin renewal and maintenance. Further exploration of Kallikrein-5's functions may deepen our understanding of its role in skin biology and its potential implications in various physiological contexts.

Measured by its ability to cleave the fluorogenic peptide substrate BOC-Val-Pro-Arg-AMC(HY-137784)that incubate at room temperature in kinetic mode for 5 minutes.The specific activity is ≥220 pmol/min/µg.

Materials
Assay buffer: 100 mM NaH₂PO₄, pH 8.0
Test Protein: Kallikrein-5 Protein, Human (HEK293, His) (HY-P70939)
Substrate: BOC-Val-Pro-Arg-AMC (HY-D1670)
Standard: 7-amino, 4-Methyl Coumarin (AMC, HY-D0027)

Procedure
1. Dilute AMC in assay buffer to concentrations of 100, 50, 25, 12.5, 6.25, 3.125, 1.5625, and 0 μmol/L. Add 100 μL of each standard concentration to a 96-well plate. Measure fluorescence in kinetic mode for 5 minutes at excitation and emission wavelengths of 380 nm and 460 nm, respectively. Plot the standard concentrations on the x-axis and the measured RFU values on the y-axis to generate a standard curve and obtain the curve equation.
2. Dilute the incubated target protein to 2 μg/mL in assay buffer.
3. Dilute the substrate to 200 μM in assay buffer.
4. Add 50 μL of the target protein to the 96-well plate, then add 50 μL of 200 μM substrate to initiate the reaction. For blank wells, add 50 μL of assay buffer and 50 μL of 200 μM substrate.
5. Read in kinetic mode for 5 minutes at excitation and emission wavelengths of 380 nm and 460 nm, respectively.
6. Calculate Specific Activity:

Human

HEK293

C-6*His

Q9Y337 (V23-S293)

25818  [NCBI]

Full Length of Mature Protein

VTEHVLANNDVSCDHPSNTVPSGSNQDLGAGAGEDARSDDSSSRIINGSDCDMHTQPWQAALLLRPNQLYCGAVLVHPQWLLTAAHCRKKVFRVRLGHYSLSPVYESGQQMFQGVKSIPHPGYSHPGHSNDLMLIKLNRRIRPTKDVRPINVSSHCPSAGTKCLVSGWGTTKSPQVHFPKVLQCLNISVLSQKRCEDAYPRQIDDTMFCAGDKAGRDSCQGDSGGPVVCNGSLQGLVSWGDYPCARPNRPGVYTNLCKFTKWIQETIQANS

Approximately 35-44 kDa

• 
  ≥ 95%, as determined by reducing SDS-PAGE.

Solution

Supplied as a 0.2 μm filtered solution of 20 mM MES, 150 mM NaCl, 10% Glycerol, pH 5.5.

<1 EU/μg, determined by LAL method.

N/A.

Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping with dry ice.