Human Serpin E1/PAI-1 Antibody

Catalog # Availability Size / Price Qty
AF1786
AF1786-SP
Detection  of Human Serpin E1/PAI‑1 by Western Blot.
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Human Serpin E1/PAI-1 Antibody Summary

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

Western Blot Detection  of Human Serpin E1/PAI‑1 by Western Blot. View Larger

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. View Larger

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.

Simple Western Detection  of Human Serpin E1/PAI‑1 by Simple  WesternTM. View Larger

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 Neutralization of Serpin E1/ PAI‑1 Activity by Human Serpin E1/PAI‑1 Antibody. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

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.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

Product Datasheets

Citations for Human Serpin E1/PAI-1 Antibody

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.

2 Citations: Showing 1 - 2
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  1. An essential role for Wnt/?-catenin signaling in mediating hypertensive heart disease
    Authors: Y Zhao, C Wang, C Wang, X Hong, J Miao, Y Liao, L Zhou, Y Liu
    Sci Rep, 2018;8(1):8996.
    Species: Rat
    Sample Types: Tissue Homogenates
    Applications: WB
  2. 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 Types: Whole Tissue
    Applications: IHC Frozen

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