Serpin E1 Protein, Mouse (HEK293, His)

Serpin E1 protein is a serine protease inhibitor that regulates TMPRSS7 and is a major inhibitor of tissue-type plasminogen activator (PLAT) and urokinase-type plasminogen activator (PLAU).It can significantly downregulate fibrinolysis and control thrombus degradation.Serpin E1 Protein, Mouse (HEK293, His) is the recombinant mouse-derived Serpin E1 protein, expressed by HEK293 , with C-6*His labeled tag.

Serpin E1, a serine protease inhibitor, exerts its regulatory functions through various mechanisms. It inhibits TMPRSS7 and serves as a primary inhibitor for both tissue-type plasminogen activator (PLAT) and urokinase-type plasminogen activator (PLAU). Acting as a crucial PLAT inhibitor, it plays a vital role in down-regulating fibrinolysis and orchestrates the controlled degradation of blood clots. In its capacity as a PLAU inhibitor, Serpin E1 is implicated in the regulation of cell adhesion and spreading. Beyond its role as a protease inhibitor, it acts as a regulator of cell migration, notably contributing to the stimulation of keratinocyte migration during cutaneous injury repair. Furthermore, Serpin E1 is involved in cellular and replicative senescence, playing a role in alveolar type 2 cells' senescence in the lung. Additionally, it participates in the regulation of cementogenic differentiation of periodontal ligament stem cells and influences odontoblast differentiation and dentin formation during odontogenesis. Serpin E1 forms a heterodimer with TMPRSS7, interacts with various proteins like VTN, PPP1CB, SORL1, LRP1, and PLAUR, and engages in complex interactions in different cellular contexts, emphasizing its multifaceted regulatory role.

Measured by its ability to inhibit uPA cleavage of 200µM N-carbobenzyloxy-Gly-Gly-Arg-7-amido-4-methylcoumarin(Z-GGR-AMC) that incubate at room temperature for 15minutes.The IC₅₀ value is 6.138 nM.

Materials
Assay buffer: 50 mM Tris, 0.01% (v/v) Tween? 20, pH 8.5
Serpin E1 Protein, Mouse (HEK293, His) (HY-P71133)
PLAU/uPA Protein, Human (411a.a, HEK293, His, solution) (HY-P71050)
Substrate: Z-Gly-Gly-Arg-AMC acetate (HY-P0019A)

Procedure
1. Dilute Human uPA to 2 μg/mL with assay buffer.
2. Prepare different concentrations of Human Serpin E1 with assay buffer for the following serial dilutions: 1500, 750, 375, 187.5, 125, 75, 50, 25, and 12.5 nM.
3. Equal volumes of Human Serpin E1/PAI-1 curves were mixed with diluted Human uPA at 2 μg/mL. Two controls containing only Human uPA and assay buffer were included.
4. The above mixture was incubated 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 , read in kinetic mode for 5 minutes.
9. The 50% inhibitory concentration (IC₅₀) of Human Serpin E1 was derived by plotting RFU/min (or specific activity) versus concentration using a 4-PL fit.

Mouse

HEK293

C-6*His

G5E899 (T23-P402)

18787  [NCBI]

Full Length of Mature Protein

TLPLRESHTAHQATDFGVKVFQQVVQASKDRNVVFSPYGVSSVLAMLQMTTAGKTRRQIQDAMGFKVNEKGTAHALRQLSKELMGPWNKNEISTADAIFVQRDLELVQGFMPHFFKLFQTMVKQVDFSEVERARFIINDWVERHTKGMISDLLAKGAVDELTRLVLVNALYFSGQWKTPFLEASTHQRLFHKSDGSTVSVPMMAQSNKFNYTEFTTPDGLEYDVVELPYQGDTLSMFIAAPFEKDVHLSALTNILDAELIRQWKGNMTRLPRLLILPKFSLETEVDLRGPLEKLGMPDMFSATLADFTSLSDQEQLSVAQALQKVRIEVNESGTVASSSTAFVISARMAPTEMVIDRSFLFVVRHNPTETILFMGQVMEP

Approximately 48.54 kDa

• 
  ≥ 95%, as determined by reducing SDS-PAGE.

Lyophilized powder.

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