Recombinant Human Pappalysin-1/PAPP-A (aa 82-1214), CF

Product Discontinued

Recombinant Human Pappalysin-1/PAPP-A (aa 82-1214), CF Summary

• Details of Functionality: Measured by its ability to cleave IGFBP-5. Cleavage of Recombinant Human IGFBP‑5 (Catalog # 875-B5) by Recombinant Human Pappalysin‑1/PAPP‑A is >50%, as measured under the described conditions.
  The IGFBP-5 cleaving activity of rhPAPP-A can be inhibited by 10 mM 1,10-phenanthroline.
• Source: Mouse myeloma cell line, NS0-derived human Pappalysin-1/PAPP-A protein
  Glu82-Asp1214 and Arg84-Asp1214, both with a C-terminal 10-His tag
• Accession #: Q13219
• N-terminal Sequence: Glu82 & Arg84
• Structure / Form: Disulfide-linked homodimer
• Protein/Peptide Type: Recombinant Enzymes
• Gene: PAPPA
• Purity: >90%, 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: 128 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: 115 kDa and 150 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 Tris, NaCl and CaCl₂.
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Assay Procedure:
  • Assay Buffer: 50 mM Tris, 10 mM CaCl₂, 150 mM NaCl, pH 7.5 (TCN)
  • Recombinant Human Pappalysin-1/PAPP-A (rhPappalysin-1) (Catalog # 2487-ZN)
  • Recombinant Human IGFBP‑5 (rhIGFBP-5) (Catalog # 875-B5)
  • Reducing SDS-PAGE gel buffer
  • SDS-PAGE and/or Western blot
  1. Dilute rhPappalysin-1 to 92.8 µg/mL in Assay Buffer.
  2. Dilute rhIGFBP-5 to 200 µg/mL in Assay Buffer.
  3. Combine equal volumes of 92.8 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 30 minute incubation period.
  5. Combine equal volumes of rhPappalysin-1/rhIGFBP-5 reaction mixuture 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: 0.464 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 82-1214), 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 ofPAPP-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, not IGFBP-5, by PAPP-A (5). The C-terminal LNR (residues 1476 to 1503) is not present in rhPAPP-A (residues 82 to 1214), which starts at the N-terminus of the mature chain and ends before the five Sushi (SCR) modules. As an active protease, rhPAPP-A cleaves IGFBP-5, which can be inhibited by 1,10-phenanthroline.

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-ZN)(2)

Publications using 2487-ZN

• M Mohrin, J Liu, J Zavala-Sol, S Bhargava, J Maxwell Tr, A Brito, D Hu, D Brooks, G Koukos, L Alabdulaal, JS Paw, K Hake, G Kolumam, ML Bouxsein, R Baron, Y Kutskova, A Freund Inhibition of longevity regulator PAPP-A modulates tissue homeostasis via restraint of mesenchymal stromal cells Aging Cell, 2021-02-09;0(0):e13313. 2021-02-09 [PMID: 33561324] (Bioassay, Human)
• Nishizawa H, Pryor-Koishi K, Suzuki M Increased levels of pregnancy-associated plasma protein-A2 in the serum of pre-eclamptic patients. Mol. Hum. Reprod., 2008-09-18;14(10):595-602. 2008-09-18 [PMID: 18805800] (Antibody Absorption, Human)

FAQs for Pappalysin-1/PAPP-A (2487-ZN). (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()