Recombinant Mouse Pappalysin-1/PAPP-A His-tag Protein, CF

R&D Systems | Catalog # 10174-ZN

R&D Systems
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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Pappalysin-1/PAPP-A His-tag Protein (10174-ZN)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

Applications

Enzyme Activity
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Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Pappalysin-1/PAPP-A protein
Ala81-Gly1624
with a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ala81

Predicted Molecular Mass

173 kDa

SDS-PAGE

141-200 kDa, under reducing conditions

Activity

Measured by its ability to cleave IGFBP-5.

Recombinant Mouse Pappalysin-1/PAPPA will cleave >50% of Recombinant Human IGFBP‑5 (Catalog # 875-B5), as measured under the described conditions.

Formulation, Preparation, and Storage

10174-ZN
Formulation Supplied as a 0.2 μm filtered solution in Tris, CaCl2, NaCl and CHAPS.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Background: Pappalysin-1/PAPP-A

Pappalysins belong to a fifth family of metzincins that consists of ADAMs/ADAMTSs, MMPs, astacins and serralysins (1). Pappalysin-1 (PAPP-A) is a secreted, disulfide-linked 400 kDa homodimer with an N-terminal catalytic domain and five C-terminal Sushi domains (2). PAPP-A cleaves Insulin-like Growth Factor-Binding Protein-4 and -5 (IGFBP-4 and -5) at a single site, resulting in the release of bioactive IGF (3). PAPP-A also contains three Lin12-Notch repeats (LNR) that bind calcium and are required for cleavage of IGFBP-4, but not IGFBP-5 (2). PAPP-A is thought to be the only physiological IGFBP-4 proteinase supported by the observation that no IGFBP-4 cleavage occurred in embryonic fibroblasts derived from PAPP-A knockout mice (4).  As a regulator of IGF bioavailability, and consequently IGF signaling, PAPP-A is an attractive therapeutic target (5-8).  PAPP-A knockout mice have an extended lifespan (9) and use of specific neutralizing antibodies causes inhibition of atherosclerotic plaque progression (10). PAPP-A levels are elevated in cancer (11-14) and PAPP-A has been identified as a migration/invasion-promoting gene (5,6, 12). PAPP-A is also important during pregnancy where PAPP-A levels are increased in the plasma by a factor of about 150. Low maternal serum levels of PAPP-A is used as an indication of Down syndrome or other fetal aneuploidies in the first trimester of pregnancy (15).

References

  1. Boldt, H. B. et al. (2001) Biochem. J. 358:359.
  2. Boldt, H. B. et al. (2004) J. Biol. Chem. 279:38525.
  3. Laursen, L. S. et al. (2001) FEBS Lett. 504:36.
  4. Conover, C. A. et al. (2004) Development 131:1187.
  5. Huang, J. et al. (2013) Oncotarget. 4:1172.
  6. Prithviraj, P. et al. (2015) Oncotarget 6: 15953.
  7. Yu, X. E. et al. (2018) Atherosclerosis. 278:250.
  8. Guo, Y. et al. (2018) Am. J. Cancer Res. 8:955.
  9. Conover, C. A. et al. (2010) J. Gerontol. A. Biol. Sci. Med. Sci. 65:590.
  10. Conover, C. A. et al. (2016) J Cardiovasc. Transl. Res. 9:77.
  11. Bulut, I. et al. (2009) Am. J. Med. Sci. 337: 241.
  12. Boldt, H. B. and Conover C. A. (2011) Endocrinology 152:1470.

Long Name

Pregnancy-associated Plasma Protein 1

Alternate Names

ASBABP2, DIPLA1, IGFBP-4ase, PAPA, PAPP-A1, PAPPA, Pappalysin1

Entrez Gene IDs

5069 (Human); 18491 (Mouse)

Gene Symbol

PAPPA

UniProt

Additional Pappalysin-1/PAPP-A Products

Product Documents for Recombinant Mouse Pappalysin-1/PAPP-A His-tag Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Mouse Pappalysin-1/PAPP-A His-tag Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Mouse Pappalysin-1/PAPP-A His-tag Protein, CF (10174-ZN):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl (TCN)
  • Recombinant Mouse Pappalysin-1/PAPPA (rmPappalysin-1) (Catalog # 10174-ZN)
  • Recombinant Human IGFBP‑5 (Catalog # 875-B5)
  • Reducing SDS-PAGE Gel Buffer
  • SDS-PAGE or Western Blot
  1. Dilute rmPappalysin-1 to 10 µg/mL in Assay Buffer.
  2. Dilute rhIGFBP-5 to 200 µg/mL in Assay Buffer.
  3. Combine equal volumes of 10 µg/mL of rmPappalysin-1 and 200 µg/mL rhIGFBP-5.  Include two controls containing Assay Buffer in place of rmPappalysin-1.
  4. Incubate reaction vials and one control at 37 C for 1 hour.  Keep the other control at -20 °C during the incubation period.
  5. Combine equal volumes of rmPappalysin-1/rhIGFBP-5 reaction mixture and reducing SDS-PAGE gel buffer.
  6. Analyze the cleavage by SDS-PAGE (load 20 µL/Lane) followed by protein staining and/or Western blot.

Per Lane:

  • rmPappalysin-1/PAPPA: 0.050 µg
  • rhIGFBP-5: 1 µg

FAQs

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