ACE2 蛋白, Human (HEK293, His)

The ACE2 protein is an important regulator of blood volume and cardiovascular homeostasis. ACE2 Protein, Human (HEK293, His) is the recombinant human-derived ACE2 protein, expressed by HEK293 , with C-His labeled tag.

ACE2 是肾素-血管紧张素激素系统中不可或缺的反调节羧肽酶，通过复杂地调节血容量和全身血管阻力，在维持心血管稳态中发挥着关键作用。通过其酶活性，ACE2 将血管紧张素 I 转化为血管紧张素 1-9，将血管紧张素 II 转化为血管紧张素 1-7，在心肌细胞中发挥抗肥厚作用，并作为具有抗增殖特性的血管舒张剂。除了在肾素-血管紧张素系统中发挥核心作用外，ACE2 还表现出广泛的酶活性，可裂解各种血管活性肽，例如神经降压素、运动紧张素和去精氨酸缓激肽。此外，ACE2 擅长切割其他生物肽，包括 apelins、酪啡肽和强啡肽 A。值得注意的是，ACE2 的 C 末端与collectrin 同源，协调中性氨基酸转运蛋白 SL6A19 向肠上皮细胞膜的运输，从而调节其表面表达和催化活性。重要的是，ACE2 还充当人类冠状病毒 SARS-CoV、SARS-CoV-2 和 HCoV-NL63 的受体，这表明它与微生物感染途径有关。

1.Measured by its binding ability in a functional ELISA. Immobilized 2019-nCoV Spike Protein (RBD, mFc) at 2 μg/mL (100 μL/well) can bind human ACE2 protein (His) and the EC₅₀ is 10-30 ng/mL.
2.Measured by its ability to cleave a fluorogenic peptide substrate McaYVADAPK (Dnp)OH and the specific activity is >1000 pmols/min/μg.

Materials
Detection Buffer: 50 mM Tris, 1 M NaCl, pH 7.5
ACE2 Protein, Human (HEK293, His) (HY-P73489)
Substrate: Mca-YVADAPK(Dnp)-OH (HY-P2616A), dissolved in DMSO to 2 mM
Standard: MCA-Pro-Leu-OH

Procedure
1. Dilute MCA-Pro-Leu-OH to 100, 50, 25, 12.5, 6.25, 3.125, 1.5625, 0 μmol/L with detection buffer respectively. Add 100 μL of each to the wells to start the reaction. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read) respectively. Plot a standard curve with the standard concentration as the abscissa and the measured RFU as the ordinate to obtain the standard curve formula.
2. Dilute Human ACE2 to 1 μg/mL with detection buffer.
3. Dilute the substrate to 20 μM with detection buffer.
4. Add 1 μg/mL Human ACE2 to the black well plate, 50 μL per well, and add 50 μL of 20 μM substrate. For blank wells, add 50 μL detection buffer and 50 μL of 20 μM substrate.
5. Read in kinetic mode for 5 minutes at excitation and emission wavelengths of 320 nm and 405 nm respectively.
6. Calculate the specific activity:

Human

HEK293

C-His

Q9BYF1-1 (Q18-S740)

59272  [NCBI]

Extracellular Domain

QSTIEEQAKTFLDKFNHEAEDLFYQSSLASWNYNTNITEENVQNMNNAGDKWSAFLKEQSTLAQMYPLQEIQNLTVKLQLQALQQNGSSVLSEDKSKRLNTILNTMSTIYSTGKVCNPDNPQECLLLEPGLNEIMANSLDYNERLWAWESWRSEVGKQLRPLYEEYVVLKNEMARANHYEDYGDYWRGDYEVNGVDGYDYSRGQLIEDVEHTFEEIKPLYEHLHAYVRAKLMNAYPSYISPIGCLPAHLLGDMWGRFWTNLYSLTVPFGQKPNIDVTDAMVDQAWDAQRIFKEAEKFFVSVGLPNMTQGFWENSMLTDPGNVQKAVCHPTAWDLGKGDFRILMCTKVTMDDFLTAHHEMGHIQYDMAYAAQPFLLRNGANEGFHEAVGEIMSLSAATPKHLKSIGLLSPDFQEDNETEINFLLKQALTIVGTLPFTYMLEKWRWMVFKGEIPKDQWMKKWWEMKREIVGVVEPVPHDETYCDPASLFHVSNDYSFIRYYTRTLYQFQFQEALCQAAKHEGPLHKCDISNSTEAGQKLFNMLRLGKSEPWTLALENVVGAKNMNVRPLLNYFEPLFTWLKDQNKNSFVGWSTDWSPYADQSIKVRISLKSALGDKAYEWNDNEMYLFRSSVAYAMRQYFLKVKNQMILFGEEDVRVANLKPRISFNFFVTAPKNVSDIIPRTEVEKAIRMSRSRINDAFRLNDNSLEFLGIQPTLGPPNQPPVS

The product has a predicted molecular mass of 83.60 kDa, and migrates as an approximately 110 kDa band in SDS-PAGE under reducing conditions.

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

无菌粉末

Lyophilized from a 0.2 μm filtered solution of PBS, pH 7.4.

<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.