Detects human Serpin E1/PAI‑1 in direct ELISAs and Western blots. In direct ELISAs, approximately 20% cross-reactivity with recombinant mouse Serpin E1 is observed and less than 2% cross-reactivity with recombinant human (rh) Serpin A1, rhSerpin A3, rhSerpin A4, rhSerpin A5, rhSerpin C1, rhSerpin F2, and rhSerpin F1/PEDF is observed.
Polyclonal Goat IgG
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human Serpin E1/PAI‑1 Gly21-Pro402 Accession # P05121
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Measured by its ability to neutralize Recombinant Human Serpin E1 (1.35 µg/mL, Catalog # 1786-PI) inhibition of Recombinant Human u‑Plasminogen Activator (uPA)/Urokinase (0.1 µg/mL, Catalog # 1310-SE) cleavage of the fluorogenic peptide substrate Z-GGR-AMC (100 µM). The Neutralization Dose (ND50) is typically 10 µg/mL.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human Serpin E1/PAI‑1 by Western Blot. Western blot shows lysates of HUVEC human umbilical vein endothelial cells. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Serpin E1/PAI‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1786) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Serpin E1/PAI‑1 at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Serpin E1/PAI‑1 in Human Liver Cancer Tissue. Serpin E1/PAI‑1 was detected in immersion fixed paraffin-embedded sections of human liver cancer tissue using Goat Anti-Human Serpin E1/PAI‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1786) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Serpin E1/PAI‑1 by Simple WesternTM. Simple Western lane view shows lysates of HUVEC human umbilical vein endothelial cells, loaded at 0.2 mg/mL. A specific band was detected for Serpin E1/PAI‑1 at approximately 54 kDa (as indicated) using 2.5 µg/mL of Goat Anti-Human Serpin E1/PAI‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1786) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Neutralization of Serpin E1/ PAI‑1 Activity by Human Serpin E1/PAI‑1 Antibody. Recombinant Human u‑Plasminogen Activator (uPA)/Urokinase (0.1 µg/mL, Catalog # 1310-SE) activity is measured in the presence of Recombinant Human Serpin E1 (1.35 µg/mL, Catalog # 1786-PI) that has been preincubated with increasing concentrations of Goat Anti-Human Serpin E1/PAI‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1786). The ND50 is typically 10 µg/mL.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Serpin E1/PAI-1
As a member of the Serpin superfamily of serine protease inhibitors, Serpin E1/PAI-1 is the principle inhibitor of urokinase-type plasminogen activator (uPA) and tissue-type PA (1, 2). As important regulators of extracellular matrix remodeling, uPA and PAI-1 play a major role in many processes such as angiogenesis, tumor invasion and obesity (2-4). For example, uPA and PAI-1 are the only tumor prognostic factors validated at the highest level of evidence with regard to their clinical utility in breast cancer (5). The human PAI-1 is initially synthesized as 402 amino acid precursor with a N-terminal signal peptide (6, 7). PAI-1 may exist in one of two possible conformations, designated as active or latent (8). The purified recombinant human (rh) PAI-1 is active against rhuPA. The heterogeneity at the N-terminus of the purified rhPAI-1 has been observed before for both the recombinant and native proteins (9).
Silverman, G.A. et al. (2001) J. Biol. Chem. 276:33293.
Stefansson, S. et al. (2003) Curr. Pharm. Des. 9:1545.
Duffy, M.J. (2002) Clin. Chem. 48:1194.
Juhan-Vague, I. et al. (2003) J. Thromb. Haemost. 1:1575.
Harbeck, N. et al. (2002) Clin. Breast Cancer 3:196.
Pannekoek, H. et al. (1986) EMBO J. 5:2539.
Ginsburg, D. et al. (1986) J. Clin. Invest. 78:1673.
Wang, Z. et al. (1996) Biochemistry 35:16443.
Stromqvist, M. et al. (1994) Protein Expr. Purif. 5:309.
Plasminogen Activator Inhibitor
Entrez Gene IDs:
5054 (Human); 18787 (Mouse)
Endothelial plasminogen activator inhibitor; Nexin; PAI1; PAI-1; PAI1PAI-1; PAISerpin E1; PLANH1; PLANH1plasminogen activator inhibitor 1; serine (or cysteine) proteinase inhibitor, clade E (nexin, plasminogenactivator inhibitor type 1), member 1; serpin peptidase inhibitor, clade E (nexin, plasminogen activator inhibitortype 1), member 1
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.