Kallikrein-5 Protein, Mouse (271a.a, HEK293, His)

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, Mouse (271a.a, HEK293, His) is the recombinant mouse-derived Kallikrein-5 protein, expressed by HEK293 , with C-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 400 µM 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 >1,350 pmol/min/μg. (Activation description: The proenzyme needs to be activated by Human Kallikrein-5 (HY-P70939) for an activated form.)

Materials
Assay buffer: 50 mM Tris, 0.005% (w/v) Brij-35, pH 8.0
Test protein: Kallikrein-5 Protein, Mouse (271a.a, HEK293, His) (HY-P77731A)
Activator protein: Kallikrein-5 Protein, Human (HEK293, His, solution) (HY-P70939)
Substrate: BOC-Val-Pro-Arg-AMC (HY-137784)
Substrate: BOC-Val-Pro-Arg-AMC (HY-137784)
Standard: 7-Amino, 4-Methyl Coumarin (AMC, HY-D0027)

Procedure
1. Standard curve: Dilute AMC to 0, 6.25, 12.5, 25, 50, 100 μmol/L using detection buffer. Add 100 μL of each dilution to black microplate wells. Measure at excitation and emission wavelengths of 380 nm and 460 nm (top read). Plot the measured RFU values on the x-axis against the standard substance concentration on the y-axis to construct the standard curve and derive the standard curve equation.
2. Dilute Mouse KLK5 to 200 μg/mL in assay buffer.
3. Dilute Human KLK5 to 2.02 μg/mL in assay buffer.
4. Mix equal volumes of 200 μg/mL Mouse KLK5 and 2.02 μg/mL Human KLK5. As a control, mix equal volumes of 2.02 μg/mL Human KLK5 with assay buffer.
5. Incubate at 37°C for 20 hours.
6. Dilute activated Mouse KLK5 to 1 μg/mL in assay buffer; perform the same dilution for the control.
7. Dilute the substrate to 800 μM in assay buffer.
8. Add 50 μL of 1 μg/mL Mouse KLK5 and control to the plate. Add 50 μL of 800 μM substrate to all wells to initiate the reaction.
9. Read for 5 minutes in kinetic mode at excitation wavelength 380 nm and emission wavelength 460 nm (top reading).
10. Calculate specific activity:

Mouse

HEK293

C-10*His

Q9D140 (V23-N293)

68668

Partial

VSEPVLAGDVSSCDNPSGTEPSGTNRDLSTDSKSGEDTRSDSSSRIVNGSDCQKDAQPWQGALLLGPNKLYCGAVLISPQWLLTAAHCRKPVFRIRLGHHSMSPVYESGQQMFQGIKSIPHPGYSHPGHSNDLMLIKMNRKIRDSHSVKPVEIACDCATEGTRCMVSGWGTTSSSHNNFPKVLQCLNITVLSEERCKNSYPGQIDKTMFCAGDEEGRDSCQGDSGGPVVCNGKLQGLVSWGDFPCAQRNRPGVYTNLCEFVKWIKDTMNSN

Approximately 35-50 kDa due to the glycosylation.

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

Solution.

Supplied as a 0.22 μm filtered solution of 20 mM NaAC, 150 mM NaCl, pH 5.0, 10% Glycerol.

<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.