Kallikrein-5 Protein, Mouse (HEK293, His)
Based on 1 Customer Validation
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.
- Species: Mouse
- Source: HEK293
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Storage: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.
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Biological Activity
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Technical Parameters
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Product Properties
- Documentation
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Biological Activity
Description
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.
Background
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.
Verified Bioactivity
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.)
Assay Procedure
Materials
Assay buffer: 50 mM Tris, 0.005% (w/v) Brij-35, pH 8.0
Test protein: Kallikrein-5 Protein, Mouse (HEK293, His) (HY-P77731)
Activator protein: 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:
Specific Activity (pmol/min/μg) = | Adjusted Vmax* (RFU/min) x Conversion Factor ** (pmol/RFU) |
| amount of enzyme (μg) |
*Adjusted for Control
**Derived using calibration standard AMC.
Technical Parameters
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Species Mouse
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Source HEK293
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Tag C-8*His
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Accession
Q9D140 (G30-N293)
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Molecular Construction
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N-term
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Kallikrein-5 (G30-N293)
Accession # Q9D140 -
8*His
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C-term
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Protein Length
Full Length of Mature Protein
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Synonyms
KLK5; KLKL2; Kallikrein Related Peptidase 5; Kallikrein-Related Peptidase 5 Transcript Variant 5; KLK-L2; Kallikrein-Related Peptidase 5 Transcript Variant 6; SCTE; Kallikrein 5 Isoform 3 Preproprotein; Stratum Corneum Tryptic Enzyme; Kallikrein 5, Isofor
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AA Sequence
GDVSSCDNPSGTEPSGTNRDLSTDSKSGEDTRSDSSSRIVNGSDCQKDAQPWQGALLLGPNKLYCGAVLISPQWLLTAAHCRKPVFRIRLGHHSMSPVYESGQQMFQGIKSIPHPGYSHPGHSNDLMLIKMNRKIRDSHSVKPVEIACDCATEGTRCMVSGWGTTSSSHNNFPKVLQCLNITVLSEERCKNSYPGQIDKTMFCAGDEEGRDSCQGDSGGPVVCNGKLQGLVSWGDFPCAQRNRPGVYTNLCEFVKWIKDTMNSN
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Predicted Molecular Mass
29.9 kDa
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Molecular Weight
Approximately 40-50 kDa, based on SDS-PAGE under reducing conditions, due to the glycosylation.
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Purity
≥ 95%, as determined by Bis-Tris PAGE.
Product Properties
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.
Documentation
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Data Sheet (237 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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Handling Instructions (2659 KB)
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
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