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

Catalog # Availability Size / Price Qty
2487-ZNF-020

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

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to cleave IGFBP-5. Recombinant Human Pappalysin‑1/PAPP‑A will cleave >50% 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 #
N-terminal Sequence
Analysis
Glu82 & Arg84
Predicted Molecular Mass
173 kDa
SDS-PAGE
139-238 kDa, three bands, reducing conditions

Product Datasheets

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2487-ZNF

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

2487-ZNF

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, CaCl2, 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.

Assay Procedure

Materials
  • 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
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Background: Pappalysin-1/PAPP-A

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).

References
  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.
Long Name
Pregnancy-associated Plasma Protein 1
Entrez Gene IDs
5069 (Human); 18491 (Mouse)
Alternate Names
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

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

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. Pregnancy-Associated Plasma Protein A Induces Inflammatory Cytokine Expression by Activating IGF-I/PI3K/Akt Pathways
    Authors: W Li, H Li, L Zhou, Z Wang, B Hua
    Mediators Inflamm., 2019;2019(0):8436985.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Cell Culture
  2. EWS-FLI-1 creates a cell surface microenvironment conducive to IGF signaling by inducing pappalysin-1
    Authors: P Jayabal, PJ Houghton, Y Shiio
    Genes Cancer, 2017;8(11):762-770.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay

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