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

• Reactivity: Mu
• Applications: Enzyme Activity
• Format: Carrier-Free

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

• Details of Functionality: 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.

• Source: Mouse myeloma cell line, NS0-derived mouse Pappalysin-1/PAPP-A protein
  Ala81-Gly1624
  with a C-terminal 6-His tag
• Accession #: Q8R4K8-1
• N-terminal Sequence: Ala81
• Protein/Peptide Type: Recombinant Enzymes
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 173 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: 141-200 kDa, under 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 Tris, CaCl₂, NaCl and CHAPS.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Assay Procedure:
  • Assay Buffer: 50 mM Tris, 10 mM CaC_l2, 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

Notes

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

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

• ASBABP2
• DIPLA1
• EC 3.4.24.79
• IGFBP-4ase
• IGFBP-4aseaspecific BCL2 ARE-binding protein 2
• IGF-dependent IGFBP-4 protease
• insulin-like growth factor-dependent IGF binding protein-4 protease
• Insulin-like growth factor-dependent IGF-binding protein 4 protease
• PAPA
• PAPPA
• PAPPA1
• PAPP-A1
• PAPP-Adifferentially placenta 1 expressed protein
• Pappalysin1
• Pappalysin-1
• pregnacy-associated plasma protein A
• Pregnancy-associated plasma protein A
• pregnancy-associated plasma protein A, pappalysin 1

Background

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).

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.

FAQs for Pappalysin-1/PAPP-A (10174-ZN). (Showing 1 - 1 of 1 FAQ).

1. I am highly interested to use couple of Anti-PAPP-A antibodies from your reputed companies for my research work. I have got the information regarding the immunogen used to generate these antibodies, however could not get any information regarding the specific amino acids/epitope to which your Anti-PAPP-A Antibodies binds with the antigen (PAPP-A).
   • Unfortunately, none of our PAPP A antibodies have been epitope mapped. Many of them have also been generated against full-length PAPP A, so there are many potential binding sites.

Bioinformatics

• Uniprot:
  • Q8R4K8-1()