Carboxypeptidase M/CPM Protein, Human (HEK293, His)
Based on 1 Customer Validation
Carboxypeptidase M Protein, Human (HEK293, His) is an approximately 55.0 kDa Carboxypeptidase M protein with a His-flag. Carboxypeptidase M is a membrane-bound arginine/lysine carboxypeptidase enzyme belongs to the carboxypeptidases family.
- Species: Human
- Source: HEK293
-
Storage: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.
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
-
Biological Activity
-
Technical Parameters
-
Product Properties
- Documentation
- References
- Help & FAQs
Biological Activity
Description
Carboxypeptidase M Protein, Human (HEK293, His) is an approximately 55.0 kDa Carboxypeptidase M protein with a His-flag. Carboxypeptidase M is a membrane-bound arginine/lysine carboxypeptidase enzyme belongs to the carboxypeptidases family[1].
Background
Carboxypeptidase M is a membrane-bound arginine/lysine carboxypeptidase enzyme, cleaves C-terminal basic amino acids with a neutral pH optimum[1]. Carboxypeptidase M belongs to the carboxypeptidases family. Carboxypeptidase M is widely distributed in a variety of tissues and cells, and it constitutively expresses in an active form at the surface of specialized cells and tissues in the human body[2]. Carboxypeptidase M plays an important role in the physiological processes of blood coagulation/fibrinolysis, inflammation, food digestion and pro-hormone and neuropeptide processing[3].
Verified Bioactivity
Measured by its ability to release L-arginine from Benzoyl-Ala-Arg, with detection of the arginine amino group by o-phthaldialdehyde. The specific activity is 187829.19 pmol/min/μg that incubate for 10 minutes at room temperature.
MCE Validation Data
-
Purity - SDS-PAGE
Purity - SDS-PAGE
Assay Procedure
Materials
Assay buffer: 50 mM MES, 5 mM CaCl2, 0.2% Triton X-100, pH 6.0
Carboxypeptidase M/CPM Protein, Human (HEK293, His) (HY-P7737)
Substrate: Bz-Ala-Arg-OH (HY-P4522), 50 mM in DMSO
Standard: L-Arginine
NaOH, dissolved in distilled water to 2 M
o-Phthaldialdehyde (OPA), dissolved in DMSO to 0.373 M
2-Mercaptoethanol (HY-Y0326)
Procedure
1. Dissolve standards in assay buffer to concentrations of 31250, 15625, 7812, 3906, 1953, 976, 488, 244, 122, 61, 30.5, 15.25, and 0 pmoL. Add an equal volume of 0.2 M NaOH solution containing 15 mM OPA (with 0.1% (v/v) 2-mercaptoethanol) to each concentration gradient. Incubate at room temperature for 10 minutes, then read at excitation wavelength 330 nm and emission wavelength 450 nm.
2. Dilute the Human CPM to 0.04 μg/mL using assay buffer.
3. Dilute the substrate to 1 mM using assay buffer.
4. Mix 150 μL of 0.04 μg/mL test protein with 150 μL of 1 mM substrate to achieve final concentrations of 0.02 μg/mL and 500 μM, respectively. For the control group, add only 150 μL of 1 mM substrate.
5. Incubate at room temperature for 10 minutes.
6. Stop the reaction by adding 300 μL of 0.2 M NaOH solution containing 15 mM OPA (with 0.1% (v/v) 2-mercaptoethanol).
7. After stopping the reaction, add 150 μL of 0.04 μg/mL Human CPM to the control group.
8. Incubate at room temperature for 10 minutes.
9. Add 200 μL of the incubated sample to the plate.
10. Read in endpoint mode at excitation and emission wavelengths of 330 nm and 450 nm.
11. Calculate specific activity:
Specific Activity (pmol/min/μg) = | Adjusted Fluorescence* (RFU) x Conversion Factor ** (pmol/RFU) |
| Incubation time (min) x amount of enzyme (μg) |
*Adjusted for Substrate Blank
**Derived using calibration standard
Per Well:
Human CPM: 0.002 μg
Substrate: 250 μM
OPA: 7.5 mM
Technical Parameters
-
Species Human
-
Source HEK293
-
Tag C-6*His
-
Accession
P14384 (L18-D421)
-
Molecular Construction
-
N-term
-
CPM (L18-D421)
Accession # P14384 -
6*His
-
C-term
-
-
Protein Length
Full Length of Carboxypeptidase MChain
-
Synonyms
CPM; Urinary Carboxypeptidase B; Carboxypeptidase M; Renal Carboxypeptidase
-
AA Sequence
LDFNYHRQEGMEAFLKTVAQNYSSVTHLHSIGKSVKGRNLWVLVVGRFPKEHRIGIPEFKYVANMHGDETVGRELLLHLIDYLVTSDGKDPEITNLINSTRIHIMPSMNPDGFEAVKKPDCYYSIGRENYNQYDLNRNFPDAFEYNNVSRQPETVAVMKWLKTETFVLSANLHGGALVASYPFDNGVQATGALYSRSLTPDDDVFQYLAHTYASRNPNMKKGDECKNKMNFPNGVTNGYSWYPLQGGMQDYNYIWAQCFEITLELSCCKYPREEKLPSFWNNNKASLIEYIKQVHLGVKGQVFDQNGNPLPNVIVEVQDRKHICPYRTNKYGEYYLLLLPGSYIINVTVPGHDPHITKVIIPEKSQNFSALKKDILLPFQGQLDSIPVSNPSCPMIPLYRNLPD
-
Predicted Molecular Mass
47.3 kDa
-
Molecular Weight
Approximately 50-58 kDa, based on SDS-PAGE under reducing conditions.
-
Glycosylation
Yes
-
Purity
≥ 95%, as determined by reducing SDS-PAGE.
Product Properties
Solution
Supplied as a 0.22 μm filtered solution of 20 mM Tris-HCl, 150 mM NaCl, 1 mM ZnCl2, pH7.5.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.
<1 EU/μg, determined by LAL method.
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.
Documentation
-
Data Sheet (263 KB)
-
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)
-
Handling Instructions (2659 KB)
References
[1]. A Nagae, et al. Carboxypeptidase M in brain and peripheral nerves. J Neurochem [Content Brief]
[2]. Kathleen Deiteren, et al. Carboxypeptidase M: Multiple alliances and unknown partners. Clin Chim Acta. 2009 Jan;399(1-2):24-39. [Content Brief]
[3]. David Reverter, Crystal structure of human carboxypeptidase M, a membrane-bound enzyme that regulates peptide hormone activity. J Mol Biol [Content Brief]
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Powered by Bioz
