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

R&D Systems | Catalog # 2487-ZNF

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Pappalysin-1/PAPP-A (aa 82-1627) (2487-ZNF)
  • 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

Q13219

Applications

Enzyme Activity
View all Pappalysin-1/PAPP-A Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Pappalysin-1/PAPP-A protein
Glu82-Gly1627 (S1224Y), with a C-terminal 6-His tag

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.

N-terminal Sequence Analysis

Glu82 & Arg84

Predicted Molecular Mass

173 kDa

SDS-PAGE

139-238 kDa, three bands, reducing conditions

Activity

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.

Formulation, Preparation, and Storage

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.

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 # http://www.rndsystems.com/product_results.aspx?k=2487‑ZN">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

Alternate Names

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

Entrez Gene IDs

5069 (Human); 18491 (Mouse)

Gene Symbol

PAPPA

UniProt

Q13219

Additional Pappalysin-1/PAPP-A Products

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

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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

For research use only

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

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Protocols

View specific protocols for Recombinant Human Pappalysin-1/PAPP-A (aa 82-1627), CF (2487-ZNF):

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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