Kallikrein-5 Protein, Mouse (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 (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 the fluorogenic peptide 400 µM substrate BOC-Val-Pro-Arg-AMC (HY-137784) that incubate at room temperature in kinetic mode for 5 minutes.The specific activity is >1350 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
Kallikrein-5 Protein, Mouse (HEK293, His) (HY-P77731)
Kallikrein-5 Protein, Human (HEK293, His) (HY-P70939)
Substrate：BOC-Val-Pro-Arg-AMC (HY-137784)
Standard: 7-amino, 4-Methyl Coumarin (AMC, HY-D0027)

Procedure
1. Dilute AMC in assay buffer to concentrations of 0, 6.25, 12.5, 25, 50, and 100 μM. Add 100 μL of each dilution to a black microplate well.
2. Dilute Mouse Kallikrein-5 to 200 μg/mL in assay buffer.
3. Dilute Human Kallikrein-5 to 2.02 μg/mL in assay buffer.
4. Mix equal volumes of 200 μg/mL Mouse KLK5 and 2.02 μg/mL Human Kallikrein-5. For the control, mix equal volumes of 2.02 μg/mL Human Kallikrein-5 and assay buffer.
5. Incubate at 37°C for 20 hours.
6. Dilute activated Mouse Kallikrein-5 to 1 μg/mL in assay buffer. Perform the same dilution on the control.
7. Dilute the substrate to 800 μM in the assay buffer.
8. Add 50 μL of 1 μg/mL Mouse Kallikrein-5 and the control to the plate, then add 50 μL of 800 μM substrate to all wells to initiate the reaction.
9. Read in kinetic mode for 5 minutes at excitation wavelength 380 nm and emission wavelength 460 nm.
10. Calculate specific activity:

Mouse

HEK293

C-8*His

Q9D140 (G30-N293)

68668  [NCBI]

Full Length of Mature Protein

GDVSSCDNPSGTEPSGTNRDLSTDSKSGEDTRSDSSSRIVNGSDCQKDAQPWQGALLLGPNKLYCGAVLISPQWLLTAAHCRKPVFRIRLGHHSMSPVYESGQQMFQGIKSIPHPGYSHPGHSNDLMLIKMNRKIRDSHSVKPVEIACDCATEGTRCMVSGWGTTSSSHNNFPKVLQCLNITVLSEERCKNSYPGQIDKTMFCAGDEEGRDSCQGDSGGPVVCNGKLQGLVSWGDFPCAQRNRPGVYTNLCEFVKWIKDTMNSN

40-50 kDa

Greater than 95% as determined by Tris-Bis PAGE.

Lyophilized powder.

Lyophilized from a 0.22 μm filtered solution of 20 mM NaAc, 150 mM NaCl, pH 5.0, 8% trehalose.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in 20 mM NaAc, 150 mM NaCl (pH 5.0).

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.