ACE2 蛋白, Mouse (HEK293, His)

ACE2 protein is an important carboxypeptidase in the renin-angiotensin system, which can regulate blood volume and vascular resistance and maintain cardiovascular homeostasis. It converts angiotensin I to the antihypertrophic angiotensin 1-9 and angiotensin II to the vasodilatory angiotensin 1-7, thereby opposing vasoconstriction. ACE2 Protein, Mouse (HEK293, His) is the recombinant mouse-derived ACE2 protein, expressed by HEK293, with C-His labeled tag.

血管紧张素转换酶 ACE2 是一种金属肽酶，作为肾素-血管紧张素系统的重要反调节羧肽酶，通过将血管紧张素 I 转化为抗心肌肥大的血管紧张素 1-9、将血管紧张素 II 转化为具有血管舒张和抗增殖特性的血管紧张素 1-7，抵消血管紧张素 II 的收缩作用，调节血容量、全身血管阻力及心血管稳态，同时作为氨基酸转运蛋白 SLC6A19 的结合伴侣调控其细胞表面运输与活性，参与氨基酸转运，并可调节神经降压素、运动紧张素等多种血管活性肽及 apelins、酪啡肽等生物肽，是心血管疾病和氨基酸稳态的关键参与者；此外，ACE2 还是 SARS-CoV 和新型冠状病毒（SARS-CoV-2）的功能受体，可被 SARS-CoV 的 S1 结构域免疫沉淀并促进病毒复制，其在人类肺和小肠上皮细胞中的大量存在为病毒提供了可能的进入途径，同时也是心脏功能和血压控制的重要调节因子，可能作为 ACE1 的天然对应物发挥作用。

Measured by its ability to cleave a fluorogenic peptide substrate, Mca-YVADAPK(Dnp)-OH. The specific activity is ≤462.95 pmol/min/µg, as measured under the described conditions.

Materials
Assay buffer: 50 mM Tris, 1 M NaCl, pH 7.0
Test Protein: ACE2 Protein, Mouse (HEK293, His) (HY-P74434)
Substrate: Mca-YVADAP-Lys(Dnp)-OH TFA (HY-P2616A), dissolved in DMSO to 2 mM
Standard: MCA-Pro-Leu-OH

Procedure
1. Dilute MCA-Pro-Leu-OH with assay buffer to 50, 25, 12.5, 6.25, 3.125, 1.5625, 0.78, 0.39, and 0 μmol/L respectively. Add 100 μL of each dilution to the wells to initiate the reaction. Measure with excitation and emission wavelengths of 320 nm and 405 nm (top read) in kinetic mode for 5 minutes. Plot a standard curve using the standard concentrations as the x-axis and the measured RFU as the y-axis, and obtain the standard curve equation.
2. Dilute ACE2 to 0.5 μg/mL with assay buffer.
3. Dilute the substrate to 20 μM with assay buffer.
4. Add different concentrations of ACE2 to a black well plate at 50 μL per well, and initiate the reaction by adding 50 μL of 20 μM substrate. For blank wells, add 50 μL of assay buffer and 50 μL of 20 μM substrate.
5. Measure in kinetic mode for 5 minutes with excitation and emission wavelengths of 320 nm and 405 nm respectively.
6. Calculate the specific activity:

Mouse

HEK293

C-His

Q8R0I0-1 (Q18-T740)

70008  [NCBI]

Extracellular Domain

QSLTEENAKTFLNNFNQEAEDLSYQSSLASWNYNTNITEENAQKMSEAAAKWSAFYEEQSKTAQSFSLQEIQTPIIKRQLQALQQSGSSALSADKNKQLNTILNTMSTIYSTGKVCNPKNPQECLLLEPGLDEIMATSTDYNSRLWAWEGWRAEVGKQLRPLYEEYVVLKNEMARANNYNDYGDYWRGDYEAEGADGYNYNRNQLIEDVERTFAEIKPLYEHLHAYVRRKLMDTYPSYISPTGCLPAHLLGDMWGRFWTNLYPLTVPFAQKPNIDVTDAMMNQGWDAERIFQEAEKFFVSVGLPHMTQGFWANSMLTEPADGRKVVCHPTAWDLGHGDFRIKMCTKVTMDNFLTAHHEMGHIQYDMAYARQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLPSDFQEDSETEINFLLKQALTIVGTLPFTYMLEKWRWMVFRGEIPKEQWMKKWWEMKREIVGVVEPLPHDETYCDPASLFHVSNDYSFIRYYTRTIYQFQFQEALCQAAKYNGSLHKCDISNSTEAGQKLLKMLSLGNSEPWTKALENVVGARNMDVKPLLNYFQPLFDWLKEQNRNSFVGWNTEWSPYADQSIKVRISLKSALGANAYEWTNNEMFLFRSSVAYAMRKYFSIIKNQTVPFLEEDVRVSDLKPRVSFYFFVTSPQNVSDVIPRSEVEDAIRMSRGRINDVFGLNDNSLEFLGIHPTLEPPYQPPVT

85-110 kDa

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

无菌粉末

Lyophilized from a 0.22 μm filtered solution of 25 mM Tris-HCl, 150 mM NaCl, 1 mM ZnCl₂, pH 7.4 or PBS, pH 7.4.
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. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

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.

• [1]. Hamming I, et, al. Tissue distribution of ACE2 protein, the functional receptor for SARS coronavirus. A first step in understanding SARS pathogenesis. J Pathol. 2004 Jun;203(2):631-7.  [Content Brief]

  [2]. Yang J, et, al. Molecular interaction and inhibition of SARS-CoV-2 binding to the ACE2 receptor. Nat Commun. 2020 Sep 11;11(1):4541.  [Content Brief]