Recombinant Human Serpin B2/PAI-2 Protein, CF

Images

 
There are currently no images for SerpinB2 (9206-PI).

Every product we sell is backed by Novus' 100% Guarantee. If you have used this product, please submit your images and reviews to earn reward points.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human Serpin B2/PAI-2 Protein, CF Summary

Details of Functionality
Measured by its ability to inhibit uPA cleavage of a peptide substrate, N-carbobenzyloxy-Gly-Gly-Arg-7-amido-4-methylcoumarin (Z-GGR-AMC).

The inhibition IC50 is <15 nM, as measured under the described conditions.

Source
E. coli-derived human Serpin B2 protein
Glu2-Ala65/Ala99-Pro415, with an N-terminal Met and 6-His tag
Accession #
N-terminal Sequence
Met
Protein/Peptide Type
Recombinant Enzymes
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
44 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.
SDS-PAGE
41 kDa, reducing conditions

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Sodium Acetate, NaCl and Chaps.
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • Assay Buffer: 50 mM Tris, 0.01% Tween® 20, pH 8.5
  • Recombinant Human Serpin B2 (rhSerpin B2) (Catalog # 9206-PI)
  • Recombinant Human u-Plasminogen Activator (uPA)/Urokinase (rhuPA) (Catalog # 1310-SE)
  • Substrate: Z-Gly-Gly-Arg-AMC (Bachem, Catalog # I-1140), 10 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: Gemini EM by Molecular Devices) or equivalent
  1. Prepare a curve of rhSerpin B2 (MW: 43.8 kDa) in Assay Buffer. Make the following serial dilutions: 400, 200, 100, 50, 25, 12.5, 6.25, 3.125, and 0.3125 nM. 
  2. Dilute rhuPA to 2 μg/mL in Assay Buffer.
  3. Combine equal volumes of each point of the rhSerpin B2 curve with 2 µg/mL rhuPA. Include an enzyme control containing equal volumes of Assay Buffer and rhuPA.
  4. Incubate reaction mixtures at room temperature for 15 minutes.
  5. Dilute Substrate to 200 µM in Assay Buffer.
  6. Load 50 µL of the diluted incubated mixtures into empty wells of a plate, and start the reaction by adding 50 µL of 200 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 200 µM Substrate.
  7. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  8. Derive the 50% inhibition concentration (IC50) value for rhSerpin-B2 by plotting RFU/min (or specific activity) versus concentration with 4-PL fitting.
  9. Calculate specific activity for each point using the following formula (if needed):

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard 7-amino, 4-Methyl Coumarin (Sigma, Catalog # A9891)

Per Well:
  • rhSerpin-B2: 100, 50, 25, 12.5, 6.25, 3.125, 1.56, 0.78, and 0.078 nM
  • rhuPA: 0.050 µg
  • Substrate: 100 µM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Serpin B2/PAI-2 Protein, CF

  • HsT1201
  • Monocyte Arg-serpin
  • PAI
  • PAI2
  • PAI2Urokinase inhibitor
  • Placental plasminogen activator inhibitor
  • PLANH2
  • PLANH2PAI-2
  • plasminogen activator inhibitor 2
  • plasminogen activator inhibitor, type II (arginine-serpin)
  • serine (or cysteine) proteinase inhibitor, clade B (ovalbumin), member 2
  • Serpin B2
  • serpin peptidase inhibitor, clade B (ovalbumin), member 2
  • Urokinase Inhibitor

Background

Serpin B2, also known as PAI-2, is an approximately 60 kDa serine protease inhibitor (1). It is primarily secreted by macrophages and monocytes (2, 3) and can form disulfide-linked multimers (4, 5). Serpin B2 inhibits both the urokinase-type and tissue-type plasminogen activators (uPA and tPA) (3, 4, 6). Serpin B2 also promotes the clearance of uPA by enhancing its binding and uptake by LRP (7). It limits fibril formation by Huntington protein (HTT) and beta-Amyloid peptides (8). It promotes Th2 biased immune responses and is important for intestinal CCL2 production, monocyte recruitment, and nematode clearance (2, 9). A non-glycosylated form of Serpin B2 is retained intracellularly where it interferes with TNF-a induced apoptosis by protecting the Retinoblastoma protein (RB1) from calpain digestion (10). It also inhibits proteasome activity in activated endothelial cells (11). Serpin B2 is upregulated in HIV-1 infected PBMC, induces RB1 upregulation, and promotes HIV-1 replication (12). Human Serpin B2 shares 76% and 73% amino acid sequence identity with mouse and rat Serpin B2, respectively.
  1. Shea-Donohue, T. et al. (2014) Gut Microbes 5:254.
  2. Schroder, W.A. et al. (2010) J. Immunol. 184:2663.
  3. Ritchie, H. et al. (1999) Thomb. Haemost. 81:96.
  4. Mikus, P. et al. (1993) Eur. J. Biochem. 218:1071.
  5. Wilczynska, M. et al. (2003) EMBO J. 22:1753.
  6. Baker, M.S. et al. (1990) Cancer Res. 50:4676.
  7. Croucher, D. et al. (2006) J. Biol. Chem. 281:10206.
  8. Lee, J.A. et al. (2015) PLoS One 10:e0130136.
  9. Zhao, A. et al. (2013) J. Immunol. 190:5779.
  10. Tonnetti, L. et al. (2008) Cancer Res. 68:5648.
  11. Boncela, J. et al. (2011) J. Biol. Chem. 286:43164.
  12. Darnell, G.A. et al. (2006) J. Biol. Chem. 281:31348.

Customers Who Viewed This Item Also Viewed...

NBP1-19773
Species: Hu, Mu, Rt
Applications: WB, ELISA, ICC/IF, IHC, IHC-Fr, IHC-P
AF1310
Species: Hu
Applications: WB, IHC, IP
NBP2-20648
Species: Hu, Mu, Rt
Applications: WB, ELISA, IHC, IHC-P
NB600-930
Species: Hu, Ba
Applications: WB, ELISA
NBP1-80633
Species: Hu
Applications: ICC/IF, IHC, IHC-P
H00005555-P01
Species: Hu
NBP2-01763
Species: Hu, Mu
Applications: WB, Flow, IHC, IHC-P
NBP1-47659
Species: Hu
Applications: WB, IHC, IHC-P
H00005554-R02
Species: Hu
H00002038-M01
Species: Hu
Applications: WB, ELISA, ICC/IF
NBP1-58268
Species: Hu
Applications: WB, IHC, IHC-P
NB600-586
Species: Hu
Applications: ICC/IF, IHC, IHC-P
NBP2-34260
Species: Hu, Mu, Po, Bv, Rb
Applications: WB, Flow, ICC/IF, IHC-Fr, IHC-P, IP, PAGE
7270-IL/CF
Species: Hu
NBP2-48990
Species: Hu
Applications: ICC/IF, IHC, IHC-P
9206-PI
Species: Hu
Applications: Enzyme Activity

Publications for SerpinB2 (9206-PI) (0)

There are no publications for SerpinB2 (9206-PI).
By submitting your publication information earn gift cards and discounts for future purchases.

Reviews for SerpinB2 (9206-PI) (0)

There are no reviews for SerpinB2 (9206-PI). By submitting a review you will receive an Amazon e-Gift Card or Novus Product Discount.
  • Review with no image -- $10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen
  • Review with an image -- $25/€18/£15/$25 CAD/¥150 Yuan/¥2500 Yen
  • 1st to review with an image -- $50/€35/£30/$50 CAD/¥300 Yuan/¥5000 Yen

FAQs for SerpinB2 (9206-PI) (0)

There are no specific FAQs related to this product. Read our general customer & technical service FAQs.

Customers Who Bought This Also Bought

Contact Information

Product PDFs

Calculators

Concentration Calculator

The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.

=
÷

Review this Product

Be the first to review our Recombinant Human Serpin B2/PAI-2 Protein, CF and receive a gift card or discount.

Bioinformatics

Uniprot