Recombinant Human Pappalysin-2/PAPP-A2 Protein, CF

Catalog # Availability Size / Price Qty
1668-ZN-020
R&D Systems Recombinant Proteins and Enzymes
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Citations (3)
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Recombinant Human Pappalysin-2/PAPP-A2 Protein, CF Summary

Product Specifications

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.
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.
Source
Mouse myeloma cell line, NS0-derived human Pappalysin-2/PAPP-A2 protein
Ser234-Cys1396, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Ser234
Predicted Molecular Mass
130 kDa
SDS-PAGE
168 kDa, reducing conditions

Product Datasheets

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1668-ZN

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.

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.
  • 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, 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
  1. Dilute rhPappalysin-2 to 40 µg/mL in Assay Buffer.
  2. 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).
  3. 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.
  4. Incubate reaction vials and one control at 37 °C overnight. Keep the other control at -20 °C during the overnight incubation period.
  5. Combine 20 µL from each reaction vial with 20 µL reducing SDS-PAGE gel buffer.
  6. 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
Reconstitution Calculator

Reconstitution Calculator

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Background: Pappalysin-2/PAPP-A2

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.

References
  1. Overgaard, M.T. et al. (2001) J. Biol. Chem. 276:21849.
  2. Boldt, H.B. et al. (2001) Biochem. J. 358:359.
  3. Fialova L. and I.M. Malbohan (2002) Bratisl. Lek. Listy 103:194.
  4. Laursen, L.S. et al. (2001) FEBS Lett. 504:36.
Long Name
Pregnancy-associated Plasma Protein
Entrez Gene IDs
60676 (Human)
Alternate Names
EC 3.4.24.-; PAPPA2; PAPP-A2; PAPP-A2pappalysin-2; pappalysin 2; Pappalysin2; Pappalysin-2; PAPPE; PAPP-E; PLAC3; placenta-specific 3; pregnancy-associated plasma preproprotein-A2; Pregnancy-associated plasma protein A2; Pregnancy-associated plasma protein E1

Citations for Recombinant Human Pappalysin-2/PAPP-A2 Protein, 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.

3 Citations: Showing 1 - 3
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  1. The potential role of pregnancy-associated plasma protein-A2 in angiogenesis and development of preeclampsia
    Authors: X Chen, K Chen, Y Feng, C Ren, W Li, J Xiao, L Fan, R Beejadhurs, L Xi, S Chen
    Hypertens. Res., 2019;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  2. Expression of pregnancy-associated plasma protein A2 during pregnancy in human and mouse.
    Authors: Wang J, Qiu Q, Haider M, Bell M, Gruslin A, Christians JK
    J. Endocrinol., 2009;202(3):337-45.
    Species: N/A
    Sample Types: N/A
    Applications: Western Blot
  3. Increased levels of pregnancy-associated plasma protein-A2 in the serum of pre-eclamptic patients.
    Authors: Nishizawa H, Pryor-Koishi K, Suzuki M
    Mol. Hum. Reprod., 2008;14(10):595-602.
    Species: Human
    Sample Types: Buffer
    Applications: Western Blot

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Reviews for Recombinant Human Pappalysin-2/PAPP-A2 Protein, CF

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Recombinant Human Pappalysin-2/PAPP-A2 Protein, CF
By Anonymous on 10/21/2022
Application: Enzymatic activity in vitro
Reason for Rating: Zero enzymatic activity in a wide variety of conditions. Probably reason is that it is not the full length protein. Truncated in the middle of the SUSHI repeats required for dimerization. Do not use for anything other than as a western blot standard. ***Bio-Techne Response: Thank 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.***