Serpin E1 蛋白, Human (HEK293, His)

Serpin E1 is a serine protease inhibitor that regulates various cellular processes by inhibiting TMPRSS7, tissue-type plasminogen activator (PLAT), and urokinase-type plasminogen activator (PLAU). During fibrinolysis, it downregulates PLAT to control thrombus degradation and acts as a PLAU inhibitor to regulate cell adhesion. Serpin E1 Protein, Human (HEK293, His) is the recombinant human-derived Serpin E1 protein, expressed by HEK293 , with C-6*His labeled tag.

Serpin E1 是一种丝氨酸蛋白酶抑制剂，对多种细胞过程发挥调节作用。它抑制 TMPRSS7，并作为组织型纤溶酶原激活剂 (PLAT) 和尿激酶型纤溶酶原激活剂 (PLAU) 的主要抑制剂。在纤溶过程中，Serpin E1 对于下调 PLAT 至关重要，有助于控制血栓的降解。同时，作为 PLAU 抑制剂，它在调节细胞粘附和扩散方面发挥作用。除了作为蛋白酶抑制剂的功能外，Serpin E1 还充当细胞迁移的调节剂，影响皮肤损伤修复过程中的角质形成细胞迁移等过程。它的参与延伸到细胞和复制衰老，导致肺部 2 型肺泡细胞衰老。此外，Serpin E1参与牙周膜干细胞牙骨质形成分化的调节，并影响成牙本质细胞分化和牙本质形成。该蛋白参与各种相互作用，与 TMPRSS7 形成异二聚体，并与多个伙伴相互作用，包括 VTN、PPP1CB、PLAUR/uPAR、SORL1 和 LRP1 等，突出了其在不同细胞途径中的多方面作用。

Measured by its ability to inhibit uPA cleavage of a peptide substrate, N-carbobenzyloxy-Gly-Gly-Arg-7-amido-4-methylcoumarin (Z-GGR-AMC). The IC₅₀ value is <15 nM, as measured under the described conditions.

Materials
Assay buffer: 50 mM Tris, 0.01% (v/v) Tween? 20, pH 8.5;
Serpin E1 Protein, Human (HEK293, His) (HY-P71077)
PLAU/uPA Protein, Human (411a.a, HEK293, His, solution) (uPA, HY-P71050)
Substrate: Z-Gly-Gly-Arg-AMC (HY-P0019A)

Procedure
1. Dilute Human uPA to 2 μg/mL with assay buffer.
2. Prepare different concentrations of Serpin E1 with assay buffer by making the following serial dilutions: 1500, 750, 375, 187.5, 125, 75, 50, 25, and 12.5 nM.
3. Equal volumes of Human Serpin E/PAI-1 curves were mixed with diluted Human uPA at 2 μg/mL. Include 2 controls containing only Human uPA and assay buffer.
4. Mix the above mixture and incubate for 15 minutes at room temperature.
5. After incubation, the mixture was diluted 10-fold in assay buffer.
6. Dilute the substrate to 200 μM with assay buffer.
7. In a 96-well plate, add 50 μL of reaction mixture and start the reaction with 50 μL of 20 μM substrate.
8. Excitation and emission wavelengths 380 nm and 460 nm were read in kinetic mode for 5 minutes.
9. The 50% inhibitory concentration (IC₅₀) of Human Serpin E1 was derived by plotting RFU/min versus concentration with a 4-PL fit.

Human

HEK293

C-6*His

P05121-1 (V24-P402)

5054  [NCBI]

Full Length of Isoform-1 Mature Protein

VHHPPSYVAHLASDFGVRVFQQVAQASKDRNVVFSPYGVASVLAMLQLTTGGETQQQIQAAMGFKIDDKGMAPALRHLYKELMGPWNKDEISTTDAIFVQRDLKLVQGFMPHFFRLFRSTVKQVDFSEVERARFIINDWVKTHTKGMISNLLGKGAVDQLTRLVLVNALYFNGQWKTPFPDSSTHRRLFHKSDGSTVSVPMMAQTNKFNYTEFTTPDGHYYDILELPYHGDTLSMFIAAPYEKEVPLSALTNILSAQLISHWKGNMTRLPRLLVLPKFSLETEVDLRKPLENLGMTDMFRQFQADFTSLSDQEPLHVAQALQKVKIEVNESGTVASSSTAVIVSARMAPEEIIMDRPFLFVVRHNPTGTVLFMGQVMEP

Approximately 44-51 kDa

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

无菌粉末.

Lyophilized from a 0.2 μm filtered solution of 20 mM HAc-NaAc, 150 mM NaCl, pH 4.0 or 20 mM HAC-NaAC, 150 mM NaCl, pH 5.5.

<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.