Cathepsin B 蛋白, Rat (HEK293, His)

Cathepsin B is a lysosomal cysteine protease that plays a role in intracellular protein catabolism. Cathepsin B mediates JNK signaling pathway to regulate the migration of glioma initiation cells. Cathepsin B is involved in the pathology of chronic inflammatory diseases of the airway and joints, as well as cancer and pancreatitis. Cathepsin B Protein, Rat (HEK293, His) is the recombinant rat-derived Cathepsin B protein, expressed by HEK293 , with C-His labeled tag.

组织蛋白酶 B 是一种溶酶体半胱氨酸蛋白酶，在细胞内蛋白质分解代谢中起作用，参与其他生理过程，如免疫应答中的抗原加工、激素激活和骨转换。组织蛋白酶 B 介导 JNK 信号通路调节胶质瘤起始细胞的迁移。组织蛋白酶 B 可以增强其他蛋白酶的活性，包括基质金属蛋白酶、尿激酶 (丝氨酸蛋白酶尿激酶纤溶酶原激活剂) 和 Cathepsin D。组织蛋白酶 B 参与气道和关节慢性炎症性疾病的病理，以及癌症和胰腺炎。

Measured by its ability to cleave the fluorogenic peptide substrate Z-LR-AMC . The specific activity is >2,500 pmol/min/μg. (Activation description: The proenzyme needs to be activated in acid reducing buffer for an activated form.)

Materials
Activation buffer: 25 mM MES, 5 mM DTT, pH 5.0
Assay buffer: 25 mM MES, pH 5.0
Cathepsin B Protein, Rat (HEK293, His) (HY-P74345)
Substrate: Z-Leu-Arg-AMC (HY-142021)
Standard: 7-Amino-4-methylcoumarin (AMC, HY-D0027)

Procedure
1. Dilute AMC to 0, 0.78125, 1.5625, 3.125, 6.25, 12.5, 25, 50, and 100 μmol/L respectively with assay buffer.
2. Add 100 μL of each dilution to black microplate wells.
3. Set excitation and emission wavelengths to 380 nm and 460 nm (top read), and read in kinetic mode for 5 minutes.
4. Plot a standard curve with the measured RFU as the abscissa and the amount of standard substance as the ordinate, and obtain the standard curve equation.
5. Dilute the test protein to 10 μg/mL with activation buffer; dilute Rat Cathepsin B to 100 μg/mL with assay buffer.
6. Incubate at room temperature for 15 minutes.
7. Dilute Rat Cathepsin B to 0.2 μg/mL with assay buffer.
8. Dilute the substrate to 20 μM with assay buffer.
9. Add 50 μL of the test protein at each concentration to the assay wells, 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 without protein.
10. Set excitation and emission wavelengths to 380 nm and 460 nm (top read), and read in kinetic mode for 5 minutes.
11. Calculate the specific activity:

Rat

HEK293

C-6*His

Q6IN22 (H18-F339)

64529  [NCBI]

Full Length of Mature Protein

HDKPSFHPLSDDMINYINKQNTTWQAGRNFYNVDISYLKKLCGTVLGGPKLPERVGFSEDINLPESFDAREQWSNCPTIAQIRDQGSCGSCWAFGAVEAMSDRICIHTNGRVNVEVSAEDLLTCCGIQCGDGCNGGYPSGAWNFWTRKGLVSGGVYNSHIGCLPYTIPPCEHHVNGSRPPCTGEGDTPKCNKMCEAGYSTSYKEDKHYGYTSYSVSDSEKEIMAEIYKNGPVEGAFTVFSDFLTYKSGVYKHEAGDVMGGHAIRILGWGIENGVPYWLVANSWNVDWGDNGFFKILRGENHCGIESEIVAGIPRTQQYWGRF

Approximately 35-43 kDa due to the glycosylation.

• 
  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, 150 mM NaCl, pH 7.5. 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.

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.