Recombinant Human VAP-1 Protein, CF

Images

 
There are currently no images for VAP-1/AOC3 (3957-AO).

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 VAP-1 Protein, CF Summary

Details of Functionality
Measured by its ability to produce hydrogen peroxide during the oxidation of benzylamine. The specific activity is >8 pmol/min/μg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human VAP-1/AOC3 protein
Gly27-Asn763, with an N-terminal Met and 10-His tag, followed by a Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Ser-Ile-Glu-Gly-Arg linker
Accession #
N-terminal Sequence
Met
Structure / Form
Disulfide-linked homodimer
Protein/Peptide Type
Recombinant Enzymes
Gene
AOC3
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
85 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
80-90 kDa, reducing conditions

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in HEPES and NaCl.
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • Assay Buffer: 50 mM HEPES, pH 7.5 
  • Recombinant Human VAP‑1/AOC3 (rhVAP‑1) (Catalog # 3957-AO)
  • Coupling Enzyme: Horseradish Peroxidase (HRP) (250 - 330 U/mg) (Sigma, Catalog # P8375), 250 units/mL stock in 0.1 M Sodium Phosphate, pH 8.0
  • Substrate Component 1: Benzylamine (Sigma, Catalog # B5136), 100 mM stock in deionized water
  • Substrate Component 2: Amplex® Ultra Red (AUR) (Molecular Probes, Catalog # A36006), 10 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhVAP-1 to 20 ng/µL in Assay Buffer.
  2. Prepare the Substrate mixture 2 mM Benzylamine, 2 units/mL HRP and 100 µM AUR in Assay Buffer.
  3. In a plate, load 50 µL of 20 ng/µL rhVAP-1 and start the reaction by adding 50 µL of the Substrate mixture (step 2). Include a Substrate Blank containing 50 µL of the Assay Buffer and 50 µL of the Substrate mixture.
  4. Read at excitation and emission wavelengths of 544 nm and 590 nm (top read), respectively in kinetic mode for 5 minutes. Note: A cutoff must be set at a wavelength of 570 nm.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

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

     *Adjusted for Substrate Blank
     **Derived using a fluorescent standard prepared by incubating 50 µM AUR, 1 unit/mL HRP, 1 mM Benzylamine, and a curve of
     Hydrogen Peroxide (Sigma, Catalog # H1009) in Assay Buffer. Use this oxidized AUR curve to determine the conversion factor.

Per Well:
  • rhVAP-1: 1.0 µg
  • Benzylamine: 1 mM
  • HRP: 1 unit/mL
  • AUR: 50 µM

Notes

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

Alternate Names for Recombinant Human VAP-1 Protein, CF

  • amine oxidase, copper containing 3 (vascular adhesion protein 1)
  • AOC3
  • Copper amine oxidase
  • EC 1.4.3
  • HPAO
  • HPAOSSAO
  • Semicarbazide-sensitive amine oxidase
  • SSAO
  • VAP1
  • VAP-1
  • VAP1EC 1.4.3.21
  • VAP-1membrane primary amine oxidase
  • Vascular adhesion protein 1

Background

Vascular adhesion protein-1 (VAP-1) is a copper amine oxidase with a topaquinone cofactor. VAP-1 is a Type II integral membrane protein, but a soluble form of the enzyme is present in human serum, and its level increases in diabetes and some inflammatory liver diseases (1, 2). VAP-1 catalyzes the oxidative deamination of small primary amines such as methylamine, benzylamine, and aminoacetone in a reaction that produces an aldehyde, ammonia, and H2O2 (3). The enzyme is sensitive to inhibition by semicarbazide. VAP-1 expression is highest in the endothelium of lung, heart, and intestine, but low in tissues such as brain, spleen, kidney, and liver (4). VAP-1 vascular expression is regulated at sites of inflammation through its release from intracellular granules in which the protein is stored (5). The adhesive function of VAP-1 has been demonstrated in studies showing that the protein is important for the adherence of certain lymphocyte subtypes to inflamed endothelial tissues (6). VAP-1 mediated adhesion is involved in the process of leukocyte extravasation, an important feature of inflammatory responses. The role of VAP-1 amine oxidase activity in this process is not fully defined, but it appears to be carbohydrate-dependent (7). VAP-1 is considered to be a therapeutic target for diabetes, oxidative stress, and inflammatory diseases (8).

  1. Kurkijärvi, R. et al. (1998) J. Immunol. 161:1549.
  2. Gearing, A.J.H. and W. Newman (1993) Immunol. Today 14:506.
  3. Lizcano, J.M. et al. (1998) Biochem. J. 331:69.
  4. Smith, D.J. et al. (1998) J. Exp. Med. 188:17.
  5. Jaakkala K. et al. (2000) Am. J. Pathol. 157:463.
  6. Salmi, M. and J. Jalkanen (2001) Trends Immunol. 22:211.
  7. Salmi, M. and J. Jalkanen (1996) J. Exp. Med. 183:569.
  8. Dunkel, P. et al. (2008) Curr. Med. Chem. 15:1827.

Customers Who Viewed This Item Also Viewed...

NBP2-38868
Species: Hu
Applications: WB, ICC/IF, IHC, IHC-P
NBP1-87493
Species: Hu, Mu, Rt
Applications: WB, IHC, IHC-P
NBP1-58006
Species: Hu
Applications: WB, IHC, IHC-P
NBP2-34260
Species: Hu, Mu, Po, Bv, Rb
Applications: WB, Flow, ICC/IF, IHC-Fr, IHC-P, IP, PAGE
NB100-2527
Species: Hu, Mu, Rt
Applications: WB, Simple Western, ICC/IF, IHC, IHC-P
NBP1-81838
Species: Hu, Mu
Applications: WB, IHC, IHC-P
NBP2-24916
Species: Hu, Mu, Rt, Bv
Applications: WB, IHC, IHC-P
796-IC
Species: Mu
NBP1-49533
Species: Hu, Mu, Rt
Applications: WB, Flow, ICC/IF, IHC, IHC-P, Flow-IC
BBA16
Species: Hu
Applications: WB, IHC, IP, ELISA(Cap), ELISA(Det), ICC, ELISA(Sta)
NB100-2076
Species: Hu, Po, Ca, Fe, Gt, GP, Sh
Applications: WB, IHC, IHC-P
H00000314-D01P
Species: Hu
Applications: WB
AF3628
Species: Mu, Rt
Applications: WB, Flow, IHC, CyTOF-ready, ICC
AF1513
Species: Mu
Applications: WB, Simple Western, Flow, IHC, IP, CyTOF-ready
NBP2-56713
Species: Hu
Applications: WB, ICC/IF
NB110-41083
Species: Hu
Applications: WB, ChIP, ELISA
3957-AO
Species: Hu
Applications: Enzyme Activity

Publications for VAP-1/AOC3 (3957-AO) (0)

There are no publications for VAP-1/AOC3 (3957-AO).
By submitting your publication information earn gift cards and discounts for future purchases.

Reviews for VAP-1/AOC3 (3957-AO) (0)

There are no reviews for VAP-1/AOC3 (3957-AO). 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 VAP-1/AOC3 (3957-AO) (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 VAP-1 Protein, CF and receive a gift card or discount.

Bioinformatics

Gene Symbol AOC3
Uniprot