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PAI1 Mouse Monoclonal Antibody [A9G8]

Usd: 350

Specification

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Catalog# HA601152

PAI1 Mouse Monoclonal Antibody [A9G8]

Application

  • WB

  • IHC-P

  • IF-Tissue

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

PAI1 Mouse Monoclonal Antibody [A9G8]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Synthetic peptide within human PAI1 aa 31-80 / 402.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Tissue, FC

Molecular Weight

Predicted band size: 45 kDa

Positive Control

No heat NIH/3T3 cell lysate, no heat NIH/3T3 serum starved for 4 hours then treated with 10ng/mL TGF-β for 21 hours add 300ng/mL BFA for 18 hours cell lysate, NIH/3T3 cell lysate, NIH/3T3 serum starved for 4 hours then treated with 10ng/mL TGF-β for 21 hours add 300ng/mL BFA for 18 hours cell lysate, HUVEC cell lysate, mouse placenta tissue lysate, rat placenta tissue lysate, human breast carcinoma tissue, human placenta tissue, HepG2, NIH/3T3.

Conjugation

unconjugated

Clone Number

A9G8

RRID

AB_3071874

Reactivity Data

WB IHC-P IF-Tissue FC
Human
Tested Tested
Tested Tested
Tested Tested
Tested Tested
Mouse
Tested Tested
Not Recommended
Not Recommended
Tested Tested
Rat
Tested Tested
Not Recommended
Not Recommended

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IHC-P

  • 1:5,000

  • IF-Tissue

  • 1:1,000

  • FC

  • 1:500-1:1,000

Target

Function

Plasminogen activator inhibitor-1 (PAI-1) also known as endothelial plasminogen activator inhibitor or serpin E1 is a protein that in humans is encoded by the SERPINE1 gene. Elevated PAI-1 is a risk factor for thrombosis and atherosclerosis. PAI-1 is a serine protease inhibitor (serpin) that functions as the principal inhibitor of tissue plasminogen activator (tPA) and urokinase (uPA), the activators of plasminogen and hence fibrinolysis (the physiological breakdown of blood clots). It is a serine protease inhibitor (serpin) protein (SERPINE1). The other PAI, plasminogen activator inhibitor-2 (PAI-2) is secreted by the placenta and only present in significant amounts during pregnancy. In addition, protease nexin acts as an inhibitor of tPA and urokinase. PAI-1, however, is the main inhibitor of the plasminogen activators. PAI-1's main function entails the inhibition of urokinase plasminogen activator (uPA), an enzyme responsible for the cleavage of plasminogen to form plasmin. Plasmin mediates the degradation of the extracellular matrix either by itself or in conjunction with matrix metalloproteinases. In this scenario, PAI-1 inhibits uPA via active site binding, preventing the formation of plasmin. Additional inhibition is mediated by PAI-1 binding to the uPA/uPA receptor complex, resulting in the latter's degradation. Thus, PAI can be said to inhibit the serine proteases tPA and uPA/urokinase, and hence is an inhibitor of fibrinolysis, the physiological process that degrades blood clots. In addition, PAI-1 inhibits the activity of matrix metalloproteinases, which play a crucial role in invasion of malignant cells through the basal lamina. PAI-1 is mainly produced by the endothelium (cells lining blood vessels), but is also secreted by other tissue types, such as adipose tissue.

Background References

1. Muñoz-Galván S et al. PAI1 is a Marker of Bad Prognosis in Rectal Cancer but Predicts a Better Response to Treatment with PIM Inhibitor AZD1208. Cells. 2020 Apr

2. Cohen C et al. Glomerular endothelial cell senescence drives age-related kidney disease through PAI-1. EMBO Mol Med. 2021 Nov

Subcellular Location

Secreted.

UNIPROT

SwissProt: P05121 Human

SwissProt: P22777 Mouse

SwissProt: P20961 Rat

Synonyms

Clade E antibody

Endothelial plasminogen activator inhibitor antibody

Nexin antibody

Nexin plasminogen activator inhibitor type 1 antibody

PAI 1 antibody

PAI antibody

PAI-1 antibody

PAI1_HUMAN antibody

PLANH1 antibody

Plasminogen activator inhibitor 1 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of PAI1 on different lysates with Mouse anti-PAI1 antibody (<a href="/products/HA601152" style="font-weight: bold;text-decoration: underline;">HA601152</a>) at 1/1,000 dilution.<br /><br />Lane 1: no heat NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 2: no heat NIH/3T3 serum starved for 4 hours then treated with 10ng/mL TGF-β for 21 hours add 300ng/mL BFA for 18 hours cell lysate (20 µg/Lane)<br />Lane 3: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 4: NIH/3T3 serum starved for 4 hours then treated with 10ng/mL TGF-β for 21 hours add 300ng/mL BFA for 18 hours cell lysate (20 µg/Lane)<br />Lane 5: HUVEC cell lysate (20 µg/Lane)<br />Lane 6: Mouse placenta tissue lysate (40 µg/Lane)<br />Lane 7: Rat placenta tissue lysate (40 µg/Lane)<br /><br />Notice: no heat means the lysate is not boiled.<br /><br />Predicted band size: 45 kDa<br />Observed band size: 45/100 kDa<br /><br />Exposure time: 10 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA601152" style="font-weight: bold;text-decoration: underline;">HA601152</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/NBI02H" style="font-weight: bold;text-decoration: underline;">NBI02H</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of PAI1 on different lysates with Mouse anti-PAI1 antibody (HA601152) at 1/1,000 dilution.

    Lane 1: no heat NIH/3T3 cell lysate (20 µg/Lane)
    Lane 2: no heat NIH/3T3 serum starved for 4 hours then treated with 10ng/mL TGF-β for 21 hours add 300ng/mL BFA for 18 hours cell lysate (20 µg/Lane)
    Lane 3: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 4: NIH/3T3 serum starved for 4 hours then treated with 10ng/mL TGF-β for 21 hours add 300ng/mL BFA for 18 hours cell lysate (20 µg/Lane)
    Lane 5: HUVEC cell lysate (20 µg/Lane)
    Lane 6: Mouse placenta tissue lysate (40 µg/Lane)
    Lane 7: Rat placenta tissue lysate (40 µg/Lane)

    Notice: no heat means the lysate is not boiled.

    Predicted band size: 45 kDa
    Observed band size: 45/100 kDa

    Exposure time: 10 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601152) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (NBI02H) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Mouse anti-PAI1 antibody (<a href="/products/HA601152" style="font-weight: bold;text-decoration: underline;">HA601152</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601152" style="font-weight: bold;text-decoration: underline;">HA601152</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Mouse anti-PAI1 antibody (HA601152) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601152) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human placenta tissue with Mouse anti-PAI1 antibody (<a href="/products/HA601152" style="font-weight: bold;text-decoration: underline;">HA601152</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA601152" style="font-weight: bold;text-decoration: underline;">HA601152</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human placenta tissue with Mouse anti-PAI1 antibody (HA601152) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601152) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunofluorescence analysis of paraffin-embedded human placenta tissue labeling PAI1 with Mouse anti-PAI1 antibody (<a href="/products/HA601152" style="font-weight: bold;text-decoration: underline;">HA601152</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/HA601152" style="font-weight: bold;text-decoration: underline;">HA601152</a>, green) at 1/1,000 dilution overnight at 4 ℃, washed with PBS.<br /><br />Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

    Immunofluorescence analysis of paraffin-embedded human placenta tissue labeling PAI1 with Mouse anti-PAI1 antibody (HA601152) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA601152, green) at 1/1,000 dilution overnight at 4 ℃, washed with PBS.

    Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

  • Flow cytometric analysis of HepG2 cells labeling PAI1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA601152" style="font-weight: bold;text-decoration: underline;">HA601152</a>, 1ug/ml) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Mouse IgG Secondary antibody (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HepG2 cells labeling PAI1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA601152, 1ug/ml) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Flow cytometric analysis of NIH/3T3 cells labeling PAI1.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA601152" style="font-weight: bold;text-decoration: underline;">HA601152</a>, 1ug/ml) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Mouse IgG Secondary antibody (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of NIH/3T3 cells labeling PAI1.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA601152, 1ug/ml) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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Citation