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Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse, Rat

Applications

ELISA, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for Serpin E1/PAI-1 Antibody - BSA Free

Immunogen

A synthetic peptide made to an internal portion of the human PAI1/Serpine 1 protein (between residues 300-400) [UniProt P05121]

Reactivity Notes

Use in Rat reported in scientific literature (PMID:35386330).

Localization

Secreted.

Specificity

PAI-1 (N315) pAb detects endogenous levels of PAI-1 protein.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

45 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Serpin E1/PAI-1 Antibody - BSA Free

Western Blot: Serpin E1/PAI-1 Antibody [NBP1-19773] - Extracts from Jurkat cells.
Immunohistochemistry-Paraffin: Serpin E1/PAI-1 Antibody [NBP1-19773] - Staining of PAI1/Serpine 1 in mouse pancreas.
Western Blot: Serpin E1/PAI-1 Antibody - BSA Free [NBP1-19773] - Serpin E1/PAI-1 Antibody [NBP1-19773] - Western Blot: Serpin E1/PAI-1 Antibody [NBP1-19773] - Immunoblots showing protein expression of ALK5, pSMAD2/3, PAI-1, pVHL, andI2-actin in A498 cells after treatment with TGF-i2 at given point of time. Image collected and cropped by Citeab from the following publication (VHL status regulates transforming growth factor B signaling pathways in renal cell carcinoma. Oncotarget (2018)) licensed under a CC-BY license.

Applications for Serpin E1/PAI-1 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

5 - 10 ug/ml

Immunohistochemistry

1:50 - 1:100

Immunohistochemistry-Frozen

1:50 - 1:100

Immunohistochemistry-Paraffin

1:50 - 1:100

Western Blot

1.0 ug/ml
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 4.5 using NBP1-19773 in the following applications:

Published Applications

Read 29 publications using NBP1-19773 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Serpin E1/PAI-1

PAI-1 and PAI-2 (for plasminogen activator inhibitor-1 and -2) are members of the serpin serine proteinase inhibitor family. PAI-1 and PAI-2 have been shown to regulate uPA (urokinase-type plasminogen activator) and tPA (tissue plasminogen activator), resulting in the inhibition of proteolytic activity. Members of the serpin family generally complex with their target proteinases, then disassociate slowly into cleaved species that fold into stable inactive forms. PAI-1 can fold into the inactive state without cleavage, resulting in the latent form of PAI-1. Activity can be restored to the latent form of PAI-1 through denaturation and renaturation. PAI-2 occurs in secreted and cytosolic forms through facultative polypeptide translocation. uPA is a serine proteinase that is a member of the trypsin family. It is responsible for the cleavage of plasminogen at the Arg-Val bond to produce plasmin. uPA consists of two chains designated A and B. The A chain can be cleaved, resulting in low and high molecular mass forms of uPA.

Long Name

Plasminogen Activator Inhibitor

Alternate Names

Nexin, PAI-1, PAI1, PLANH1

Entrez Gene IDs

5054 (Human); 18787 (Mouse); 24617 (Rat)

Gene Symbol

SERPINE1

UniProt

Product Documents for Serpin E1/PAI-1 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Serpin E1/PAI-1 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Protocols

View specific protocols for Serpin E1/PAI-1 Antibody - BSA Free (NBP1-19773):

[[URL: https://www.novusbio.com/products/serpin-e1-pai-1-antibody_nbp1-19773]]… E1/PAI-1 Antibody]]
Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
[[URL:https://www.novusbio.com/products/serpin-e1-pai-1-antibody_nbp1-19773]]… E1/PAI-1 Antibody]]
Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.


[[URL:https://www.novusbio.com/products/serpin-e1-pai-1-antibody_nbp1-19773]]… E1/PAI-1 Antibody]]
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.
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