Carboxypeptidase A1/CPA1 蛋白, Mouse (HEK293, His)

Carboxypeptidase A1 (CPA1) is an enzyme responsible for catalyzing the release of a C-terminal amino acid; however, it exhibits limited or no activity with residues such as -Asp, -Glu, -Arg, -Lys, or -Pro. This specificity in substrate recognition underscores its role as a carboxypeptidase with selectivity towards certain amino acid sequences at the C-terminus of peptides. Carboxypeptidase A1/CPA1 Protein, Mouse (HEK293, His) is the recombinant mouse-derived Carboxypeptidase A1/CPA1 protein, expressed by HEK293 , with C-His labeled tag.

羧肽酶 A1 (CPA1) 是一种在催化肽 C 端氨基酸释放中发挥关键作用的酶。然而，对于具有 C 末端残基（例如 -Asp、-Glu、-Arg、-Lys 或 -Pro）的肽，其活性有限或不存在。这种底物特异性表明 CPA1 在其切割偏好方面表现出选择性，更喜欢肽 C 末端的某些氨基酸残基。该酶释放 C 末端氨基酸的作用有助于各种生理过程，例如消化系统中蛋白质的消化，凸显了其在调节肽和蛋白质代谢中的重要性。

Measured by its ability to cleave the colorimetric peptide substrate Ac-Phe-Thiaphe-OH in the presence of 5,5’Dithio-bis (2-nitrobenzoic acid) (DTNB). The specific activity is 5420.437 pmol/min/µg, as measured under the described conditions. (Activation description: The proenzyme needs to be activated by Trypsin for an activated form.)

Materials
Assay buffer: 50 mM Tris, 0.15 M NaCl, 10 mM CaCl₂, 0.05% Brij-35, pH 7.5 (TCNB)
Carboxypeptidase A1/CPA1 Protein, Mouse (HEK293, His) (HY-P72868)
Trypsin
Substrate: Ac-Phe-Thiaphe-OH
DTNB (HY-15915)

Procedure
1. Dilute Mouse CPA1 to 100 µg/mL using a detection buffer containing 1.0 µg/mL Trypsin.
2. Incubate at room temperature for 60 minutes. Dilute Mouse CPA1 to 0.2 µg/mL in the assay buffer.
3. Mix equal volumes of 10 mM substrate and 10 mM DTNB. Then dilute the mixture to 200 µM in the assay buffer.
4. Add 50 µL of 0.2 µg/mL Mouse CPA1 to a transparent microplate. Add 50 µL of assay buffer to the blank.
5. Add 50 µL of the 200 µM substrate mixture to the wells to start the reaction.
6. Read at 405 nm in kinetic mode for 5 minutes.
7. Calculate specific activity:

Mouse

HEK293

C-His

Q7TPZ8 (N17-Y419)

109697  [NCBI]

Full Length of Mature Protein

NENFVGHQVLRISATDEAQVQKVRELEELEHLKLDFWRDPARAGLPIDVRVPFPTIQSVKAFLEYHDISYEIMIEDVQSLLDEEKQQMSAFQARALSTDAFNYATYHTLDEIYEFMDLLVTEHPQLVSKIQIGSTFEGRPINVLKFSTGGTNRPAIWIDTGIHSREWVTQASGVWFAKKITKDYGQEPTLTAILDNMDIFLEIVTNPDGFVYTHKTNRMWRKTRSHTEGSLCVGVDPNRNWDAAFGMPGASSNPCSETYRGKFPNSEVEVKSIVDFVTSHGNIKAFISIHSYSQLLLYPYGYTSEPAPDKEELDQLAKSAVTALTSLHGTKFKYGSIIDTIYQASGSTIDWTYSQGIKYSFTFELRDTGLRGFLLPASQIIPTAEETWLALLTIMDHTVKHPY

Approximately 42-49 kDa

• 
  Greater than 95% as determined by reducing SDS-PAGE.

无菌粉末

Lyophilized from a 0.22 μm filtered solution of 20 mM Tris, 150 mM NaCl, pH 7.5 or 25 mM Tris-HCl, 150 mM NaCl, pH 7.5.
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.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O.

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.