Alpha-2-Antiplasmin/Serpin F2 蛋白, Rat (HEK293, His)

Alpha-2-antiplasmin/serpin F2 protein is an important member of the serine protease inhibitor family and plays a key role as a serine protease inhibitor, regulating proteolytic activity. His research enhances the understanding of cellular processes related to proteolysis and its therapeutic applications. Alpha-2-Antiplasmin/Serpin F2 Protein, Rat (HEK293, His) is the recombinant rat-derived Alpha-2-Antiplasmin/Serpin F2 protein, expressed by HEK293 , with C-His labeled tag.

Alpha-2-抗纤溶酶/丝氨酸蛋白酶抑制剂 F2 蛋白是丝氨酸蛋白酶抑制剂家族的重要成员，凸显了其作为丝氨酸蛋白酶抑制剂的关键作用。作为该家族的一部分，Alpha-2-抗纤溶酶/丝氨酸蛋白酶抑制剂 F2 可能与相关蛋白质具有保守的结构和功能特征，强调其参与调节蛋白水解活性。丝氨酸蛋白酶抑制剂家族的分类强调了其在丝氨酸蛋白酶抑制剂的更广泛背景下的特定名称，提供了对其独特机制和底物特异性的见解。Alpha-2-抗纤溶酶/丝氨酸蛋白酶抑制剂 F2 的研究有助于我们了解其在与蛋白水解相关的细胞过程中的作用，提供在治疗干预中的潜在应用，并更深入地理解其对涉及蛋白质代谢的细胞过程的更广泛影响。对 Alpha-2-抗纤溶酶/丝氨酸蛋白酶抑制剂 F2 作用的进一步探索有望增强我们对其对正常生理和病理状况的贡献的了解。

Measured by its ability to inhibit trypsin cleavage of a fluorogenic peptide substrate Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (HY-P2185A) that incubate at 37 °C for 15 minutes. The IC₅₀ value is 0.1895 nM.

Materials
Assay buffer: 50 mM Tris, 10 mM CaCl₂, 150 mM NaCl, 0.05% (w/v) Brij-35, pH 7.5 (TCNB)
Alpha-2-Antiplasmin/Serpin F2 Protein, Rat (HEK293, His) (HY-P76721)
Trypsin
Substrate: Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (NFF-3 TFA, HY-P2185A)

Procedure
1. Dilute Trypsin to 0.25 µg/mL in assay buffer and keep on ice.
2. Prepare different concentrations of Rat Serpin F2 in assay buffer with the following serial dilutions: 100, 20, 15, 10, 7, 4, 2, 1, 0 nM.
3. Mix equal volumes of Serpin F2 (various concentrations) with Trypsin (from step 1) .
4. Incubate the mixture at 37°C for 15 minutes.
5. After incubation, dilute the mixture 1/5 with assay buffer.
6. Dilute the substrate to 200 µM in assay buffer.
7. Add 50 µL of the diluted reaction mixture to the wells, followed by 50 µL of 200 µM substrate to initiate the reaction.
8. Read the fluorescence in kinetic mode for 5 minutes with excitation and emission wavelengths of 320 nm and 405 nm (top read).
9. Determine the 50% inhibitory concentration (IC₅₀) by plotting RFU/min versus concentration and fitting the data with a 4-parameter logistic (4-PL) curve.

Rat

HEK293

C-10*His

Q68FT8 (M28-K491)

287527  [NCBI]

Full Length of Mature Protein

MDLPGQQPVSEQAQQKLPPLALLKLGNQDLGDHATLKRSPGDCKSAPTTEETRRLSQAMMAFTTDLFSLVAQTSTSSNLVLSPLSVALALSHLALGARNQTLENLQRVLHMNMGSCIPHLLSHFCQNLNPGTIRLAARIYLQKGFPIKDDFLEQSEKLFGAKPVKLTGRQEEDLMNINKWVKEATEGKIEDFLSELPDNTVLLLLNAIHFHGFWRTKFDPSLTQKDSFHLDEQFTVPVAMMHAQSYPLRWFLLEQPEIQVAHFPFQNNMSFVVIMPTYFGWNVSEVLANLTWDTLYQPSMREKPTKVRLPKLHLEQHLDLVATLSKLGLQDLFQSPDLRGISDQSLVVSSVQHQSTMELSEAGVEAAAATSTAMTRMSLSSFFLNRPFIFFIMEETIGIPLFVGSVRNPDPSAQPQPQEQQDSPDNRRLDQNDKADIPGGKTFAPDLKLVPRLEEDYPQFSSPK

Approximately 58-90 kDa.

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

无菌粉末.

Lyophilized from a 0.22 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.