Cathepsin S 蛋白, Mouse (HEK293, His)

Cathepsin S Protein, Mouse (HEK293, His) is potent cysteine protease which can promote degradation of damaged or unwanted proteins in the endo-lysosomal pathway.

组织蛋白酶 S 是半胱氨酸组织蛋白酶家族的成员。组织蛋白酶 S 具有特定作用，例如 MHC II 类抗原呈递，在恒定链的降解中发挥重要作用。组织蛋白酶 S 参与多种病理过程，包括关节炎、癌症和心血管疾病，它会被分泌并作用于细胞外底物。组织蛋白酶 S 具有独特的组织表达限制，并且在中性 pH 下更稳定。组织蛋白酶 S 在半胱氨酸组织蛋白酶家族中是独特的，因为其组织表达受到限制，与位于淋巴和脾脏中的抗原呈递细胞以及巨噬细胞等其他免疫细胞相关。组织蛋白酶 S 被认为是一种特别有效的半胱氨酸蛋白酶，可裂解弹性蛋白并产生具有生物活性的弹性蛋白肽，从而促进心血管炎症和钙化。组织蛋白酶 S 也由平滑肌细胞和巨噬细胞释放，作为连续递归反馈循环中炎症的全身反应^[1][2]。

Measured by its ability to cleave the fluorogenic peptide substrate Mca-RPKPVE-Nval-WRK(Dnp)-NH2. Read at excitation and emission wavelengths of 320 nm and 405 nm. The specific activity is >300 pmol/min/µg.

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  Measured by its ability to cleave the fluorogenic peptide substrate Mca-RPKPVE-Nval-WRK(Dnp)-NH2. Read at excitation and emission wavelengths of 320 nm and 405 nm. The specific activity is 17654.2954 pmol/min/µg, as measured under the described conditions.

Materials
Assay buffer: 50 mM NaOAc,5 mM DTT, 250 mM NaCl, pH 4.5
Cathepsin S Protein, Mouse (HEK293, His) (HY-P7757)
Substrate: NFF-3 TFA (HY-P2185A)
Standard: MCA-Pro-Leu-OH

Procedure
1. Dilute the standard with the assay buffer to the following concentrations: 0, 0.5, 1, 2, 3.9, 7.8, 15.6, 31.25, 62.5, 125, 250, and 500 μmol/L. Add 100 μL of each concentration to the wells to initiate the reaction, with three replicate wells for each concentration. Read the reaction in kinetic mode for 5 minutes at excitation and emission wavelengths of 320 nm and 405 nm, respectively. Plot the standard curve with the standard concentration as the x-axis and the measured RFU as the y-axis to obtain the standard curve equation.
2. First, reconstitute the protein to a concentration of 200 μg/mL with sterile water, then dilute Mouse Cathepsin S to 10 μg/mL with the assay buffer.
3. Incubate at room temperature for 2 hours.
4. Dilute Mouse Cathepsin S to 0.5 ng/μL with the assay buffer.
5. Dilute the substrate to 20 μM in assay buffer.
6. Add 50 μL of 0.5 ng/μL Mouse Cathepsin S and 50 μL of 20 μM substrate into a 96-well black plate. Include a substrate blank containing 50 μL of assay buffer and 50 μL of 20 μM substrate without any Mouse Cathepsin S. Perform each concentration in triplicate.
7. Read the RFU values in kinetic mode at excitation and emission wavelengths of 320 nm and 405 nm, respectively, for 5 minutes.
8. Calculate the specific activity:

Mouse

HEK293

C-6*His

O70370 (V18-I340)

13040  [NCBI]

Full Length of Mature Protein (with Propeptide)

VCSVAMEQLQRDPTLDYHWDLWKKTHEKEYKDKNEEEVRRLIWEKNLKFIMIHNLEYSMGMHTYQVGMNDMGDMTNEEILCRMGALRIPRQSPKTVTFRSYSNRTLPDTVDWREKGCVTEVKYQGSCGACWAFSAVGALEGQLKLKTGKLISLSAQNLVDCSNEEKYGNKGCGGGYMTEAFQYIIDNGGIEADASYPYKATDEKCHYNSKNRAATCSRYIQLPFGDEDALKEAVATKGPVSVGIDASHSSFFFYKSGVYDDPSCTGNVNHGVLVVGYGTLDGKDYWLVKNSWGLNFGDQGYIRMARNNKNHCGIASYCSYPEIHHHHHH

Approximately 37-42 & 29-34 & 14 kDa, due to the glycosylation.

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

无菌粉末

Lyophilized from a 0.22 μm filtered solution of 20 mM PB,150 mM NaCl, pH 7.4 or 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.

• [1]. Richard D A Wilkinson, et al. Cathepsin S: therapeutic, diagnostic, and prognostic potential. Biol Chem. 2015 Aug;396(8):867-82.  [Content Brief]

  [2]. Brena F Sena, et al. Cathepsin S As an Inhibitor of Cardiovascular Inflammation and Calcification in Chronic Kidney Disease. Front Cardiovasc Med. 2018 Jan 5;4:88.  [Content Brief]