Pappalysins belong to a fifth family of metzincins that consists of ADAMs/ADAMTSs, MMPs, astacins and serrylysins (1, 2). 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 (3). 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 (4). Compared to PAPP‑A, PAPP-A2 (also called PAPP‑E), the second member of the family is less characterized. PAPP-A2 shares 45% amino acid identity to PAPP-A in the mature form, which is synthesized as a preproprotein consisting of multiple domains (1). The prepro region (residues 1‑233) and the C-terminal region (residues 1397‑1791) are not included in recombinant human PAPP-A2. As an active protease, recombinant human PAPP-A2 cleaves IGFBP-5 under the conditions described in Activity Assay Protocol.
Recombinant Human Pappalysin-2/PAPP-A2 Protein, CF
R&D Systems | Catalog # 1668-ZN
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Key Product Details
- R&D Systems NS0-derived Recombinant Human Pappalysin-2/PAPP-A2 Protein (1668-ZN)
- 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
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human Pappalysin-2/PAPP-A2 protein
Ser234-Cys1396, with a C-terminal 10-His tag
Ser234-Cys1396, with a C-terminal 10-His tag
Purity
>90%, 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
Ser234
Predicted Molecular Mass
130 kDa
SDS-PAGE
168 kDa, reducing conditions
Activity
Measured by its ability to cleave IGFBP-5.
Cleavage of Recombinant Human IGFBP‑5 (Catalog # 875-B5) by Recombinant Human Pappalysin‑2/PAPP‑A2 is detectable by SDS-PAGE stained by Coomassie, as measured under the described conditions.
Cleavage of Recombinant Human IGFBP‑5 (Catalog # 875-B5) by Recombinant Human Pappalysin‑2/PAPP‑A2 is detectable by SDS-PAGE stained by Coomassie, as measured under the described conditions.
Reviewed Applications
Read 1 review rated 1 using 1668-ZN in the following applications:
Formulation, Preparation, and Storage
1668-ZN
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and CaCl2. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: Pappalysin-2/PAPP-A2
References
- Overgaard, M.T. et al. (2001) J. Biol. Chem. 276:21849.
- Boldt, H.B. et al. (2001) Biochem. J. 358:359.
- Fialova L. and I.M. Malbohan (2002) Bratisl. Lek. Listy 103:194.
- Laursen, L.S. et al. (2001) FEBS Lett. 504:36.
Long Name
Pregnancy-associated Plasma Protein
Alternate Names
PAPP-A2, PAPPA2, Pappalysin2, PAPPE, PLAC3
Entrez Gene IDs
60676 (Human)
Gene Symbol
PAPPA2
UniProt
Additional Pappalysin-2/PAPP-A2 Products
Product Documents for Recombinant Human Pappalysin-2/PAPP-A2 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Pappalysin-2/PAPP-A2 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Pappalysin-2/PAPP-A2 Protein, CF
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Application: Enzymatic activity in vitroVerified Customer | Posted 10/21/2022Bio-Techne ResponseThank you for reviewing our product. We are sorry to hear that this product did not perform as expected. We have been in touch with the customer to resolve this issue according to our Product Guarantee and to the customer’s satisfaction. Bio-Techne Technical Service provided additional support and guidance to customer developing and performing assays which differed from product QC activity specifications.
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Protocols
View specific protocols for Recombinant Human Pappalysin-2/PAPP-A2 Protein, CF (1668-ZN):
Materials
- Assay Buffer: 50 mM Tris, pH 8.0
- Recombinant Human Pappalysin-2/PAPP-A2 (rhPappalysin-2) (Catalog # 1668-ZN)
- Recombinant Human IGFBP‑5 (rhIGFBP-5) (Catalog # 875-B5)
- 15% SDS-PAGE gel
- Reducing SDS-PAGE gel buffer
- SDS-PAGE or Western Blot
- Dilute rhPappalysin-2 to 40 µg/mL in Assay Buffer.
- Dilute rhIGFBP-5 to 200 µg/mL in Assay Buffer (Note: Acetonitrile interferes with assay, dry down rhIGFBP-5 and reconstitute at 200 µg/mL with Assay Buffer if necessary).
- Combine equal volumes of 40 µg/mL rhPappalysin-2 and 200 µg/mL rhIGFBP-5 for a 1:23.5 ratio of enzyme to Substrate. Include two blank controls containing Assay Buffer in place of rhPappalysin-2.
- Incubate reaction vials and one control at 37 °C overnight. Keep the other control at -20 °C during the overnight incubation period.
- Combine 20 µL from each reaction vial with 20 µL reducing SDS-PAGE gel buffer.
- Analyze the cleavage by SDS PAGE (load 40 µL/lane) followed by protein staining and/or Western blot.
Per Lane:
- rhPappalysin-2: 0.4 µg
- rhIGFBP-5: 2.0 µg
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