Recombinant Human Pappalysin-1/PAPP-A (aa 82-1627), CF

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

Recombinant Human Pappalysin-1/PAPP-A (aa 82-1627), CF Summary

• Details of Functionality: Measured by its ability to cleave IGFBP-5. Recombinant Human Pappalysin‑1/PAPP‑A will cleave >75% of Recombinant Human IGFBP‑5 (Catalog # 875-B5) , as measured under the described conditions.
• Source: Mouse myeloma cell line, NS0-derived human Pappalysin-1/PAPP-A protein
  Glu82-Gly1627 (S1224Y), with a C-terminal 6-His tag
• Accession #: Q13219
• N-terminal Sequence: Glu82 & Arg84
• Protein/Peptide Type: Recombinant Enzymes
• Gene: PAPPA
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Endotoxin Note: <1.0 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: 139-238 kDa, three bands, 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, NaCl, CaCl₂, and CHAPS.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Assay Procedure:
  • Assay Buffer: 50 mM Tris, 10 mM CaCl₂, 150 mM NaCl (TCN)
  • Recombinant Human Pappalysin-1/PAPP-A (rhPappalysin-1) (Catalog # 2487-ZNF)
  • Recombinant Human IGFBP‑5 (rhIGFBP‑5) (Catalog # 875-B5)
  • Reducing SDS-PAGE Gel Buffer
  • SDS-PAGE or Western Blot
  1. Dilute rhPappalysin-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 rhPappalysin-1 and 200 µg/mL rhIGFBP-5. Include two controls containing Assay Buffer in place of rhPappalysin-1.
  4. Incubate reaction vials and one control at 37 °C for 30 minutes. Keep the other control at -20 °C during the incubation period.
  5. Combine equal volumes of rhPappalysin-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:
  • rhPappalysin-1: 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 Human Pappalysin-1/PAPP-A (aa 82-1627), 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 serrylysins (1). PAPP-A is an important pregnancy protein and increases in plasma by a factor of about 150 during pregnancy as compared to the nonpregnant state. PAPP-A is also a major marker of Down syndrome in the first trimester of pregnancy because maternal serum levels of PAPP‑A are significantly reduced when a fetus affected by Down syndrome is present (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). Lack of IGFBP-4 cleavage in embryonic fibroblasts derived from PAPP-A knockout mice indicates that PAPP-A functions as a physiological IGFBP-4 protease (4). Three Lin12-Notch repeats (LNR) in the PAPP-A protein bind Ca²⁺­­­­ and are required for the cleavage of IGFBP-4, but not IGFBP-5, by PAPP-A (5). As an active protease, recombinant human (rh) PAPP-A cleaves IGFBP-4 and IGFBP-5. This full-length rhPAPP-A, which contains the five C-terminal Sushi domains, is approximately 9-fold more active for cleavage of IGFBP-5 than the C-terminally truncated rhPAPP-A (Catalog # 2487‑ZN).

1. Boldt, H.B. et al. (2001) Biochem. J. 358:359.
2. Fialova L. and I.M. Malbohan (2002) Bratisl. Lek. Listy 103:194.
3. Laursen, L.S. et al. (2001) FEBS Lett. 504:36.
4. Conover, C.A. et al. (2004) Development 131:1187.
5. Boldt, H.B. et al. (2004) J. Biol. Chem. 279:38525.

Publications for Pappalysin-1/PAPP-A (2487-ZNF)(3)

Publications using 2487-ZNF

• Hefzi, H;Martínez-Monge, I;Marin de Mas, I;Cowie, NL;Toledo, AG;Noh, SM;Karottki, KJC;Decker, M;Arnsdorf, J;Camacho-Zaragoza, JM;Kol, S;Schoffelen, S;Pristovšek, N;Hansen, AH;Miguez, AA;Bjorn, SP;Brøndum, KK;Javidi, EM;Jensen, KL;Stangl, L;Kreidl, E;Kallehauge, TB;Ley, D;Ménard, P;Petersen, HM;Sukhova, Z;Bauer, A;Casanova, E;Barron, N;Malmström, J;Nielsen, LK;Lee, GM;Kildegaard, HF;Voldborg, BG;Lewis, NE; Multiplex genome editing eliminates the Warburg Effect without impacting growth rate in mammalian cells bioRxiv : the preprint server for biology 2024-08-06 [PMID: 39211256] (In vitro assay, Bovine)
• W Li, H Li, L Zhou, Z Wang, B Hua Pregnancy-Associated Plasma Protein A Induces Inflammatory Cytokine Expression by Activating IGF-I/PI3K/Akt Pathways Mediators Inflamm., 2019-09-10;2019(0):8436985. 2019-09-10 [PMID: 31582904] (Cell Culture, Mouse)
• P Jayabal, PJ Houghton, Y Shiio EWS-FLI-1 creates a cell surface microenvironment conducive to IGF signaling by inducing pappalysin-1 Genes Cancer, 2017-11-01;8(11):762-770. 2017-11-01 [PMID: 29321818] (Bioassay, Human)

FAQs for Pappalysin-1/PAPP-A (2487-ZNF). (Showing 1 - 1 of 1 FAQs).

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

• Gene Symbol: PAPPA
• Uniprot:
  • Q13219()