Human Serpin E1/PAI-1 Antibody

(1 citations)   
  • Species Reactivity
    Human
  • Specificity
    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. 
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    S. frugiperda insect ovarian cell line Sf 21-derived recombinant human Serpin E1/PAI‑1
    Gly21-Pro402
    Accession # P05121
  • Formulation
    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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    2.5 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunoprecipitation
    25 µg/mL
    Conditioned cell culture medium spiked with Recombinant Human Serpin E1/PAI‑1 (Catalog # 1786-PI), see our available Western blot detection antibodies
  • Neutralization
    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.
Data Examples
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.
Immunohistochemistry
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
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    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 principal 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).
  • References:
    1. Silverman, G.A. et al. (2001) J. Biol. Chem. 276:33293.
    2. Stefansson, S. et al. (2003) Curr. Pharm. Des. 9:1545.
    3. Duffy, M.J. (2002) Clin. Chem. 48:1194.
    4. Juhan-Vague, I. et al. (2003) J. Thromb. Haemost. 1:1575.
    5. Harbeck, N. et al. (2002) Clin. Breast Cancer 3:196.
    6. Pannekoek, H. et al. (1986) EMBO J. 5:2539.
    7. Ginsburg, D. et al. (1986) J. Clin. Invest. 78:1673.
    8. Wang, Z. et al. (1996) Biochemistry 35:16443.
    9. Stromqvist, M. et al. (1994) Protein Expr. Purif. 5:309.  
  • Long Name:
    Plasminogen Activator Inhibitor
  • Entrez Gene IDs:
    5054 (Human); 18787 (Mouse)
  • Alternate Names:
    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 E1; serpin peptidase inhibitor, clade E (nexin, plasminogen activator inhibitortype 1), member 1
Related Research Areas
Citations:

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.

1 Citations: Showing 1 - 1

  1. Overexpression of protease nexin-1 mRNA and protein in oral squamous cell carcinomas.
    Authors: Gao S, Krogdahl A, Sorensen J, Kousted T, Dabelsteen E, Andreasen P
    Oral Oncol, 2008;44(3):309-13.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Frozen
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