Recombinant Human Notum Protein, CF

Recombinant Human Wnt-3a shows a dose responsive increase in Wnt reporter activity in HEK293 cells (green line). Recombinant Human Notum (Catalog # 9118-NM) inhibits a constant dose of 0.5 μg/mL Recombinant Human Wnt-3a (Catalog # 5036-WN). The ED50 is 0.1-0.6 µg/mL (orange line).

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human Notum Protein, CF Summary

• Details of Functionality: Measured by its ability to inhibit Wnt induced TCF reporter activity in HEK293 human embryonic kidney cells. The ED₅₀ for this effect is 0.1-0.6 μg/mL in the presence of 0.5 μg/mL of Recombinant Human Wnt-3a (Catalog # 5036-WN).
• Source: Chinese Hamster Ovary cell line, CHO-derived human Notum protein
  Ser81-Thr451 (Cys330Ser) with a C-terminal 6-His tag
• Accession #: Q6P988
• N-terminal Sequence: Ser81
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 43 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: 38-45 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Notum Protein, CF

• EC 3.-
• notum pectinacetylesterase homolog (Drosophila)
• Notum
• protein notum homolog

Background

Notum is an evolutionarily conserved 60 kDa secreted deacylase that regulates Wnt activity (1, 2). It is tethered to the cell surface by binding to Glypican-like sulfated proteoglycans, and it functions as a Glypican-dependent Wnt inhibitor (1-4). Notum hydrolyzes the palmitoyl moiety from Wnt-3a, making the Wnt more hydrophilic and unable to bind to the Frizzled-8 receptor (1, 2). It also induces the release of cell surface GPI-anchored proteins such as Glypican-3, Cadherin-13, and uPAR (4). Notum regulates head regeneration in planaria (3), head formation and neural induction in Xenopus (1), and long range activity of hedgehog proteins in Drosophila (5). It is up-regulated in some human hepatocellular carcinomas and colorectal cancers (6, 7). Within amino acids
81-451, human Notum shares 92% aa sequence identity with mouse and rat Notum.

1. Zhang, X. et al. (2015) Dev. Cell 32:719.
2. Kakugawa, S. et al. (2015) Nature 519:187.
3. Petersen, C.P. and P.W. Reddien (2011) Science 332:852.
4. Traister, A. et al. (2008) Biochem. J. 410:503.
5. Ayers, K.L. et al. (2010) Dev. Cell 18:605.
6. Torisu, Y. et al. (2008) Cancer Sci. 99:1139.
7. De Robertis, M. et al. (2015) Oncotarget 6:41237.

Publications for NOTUM (9118-NM)(1)

Publication using 9118-NM

• F Guo, M Seldin, M Péterfy, S Charugundl, Z Zhou, SD Lee, A Mouton, P Rajbhandar, W Zhang, M Pellegrini, P Tontonoz, AJ Lusis, DM Shih NOTUM promotes thermogenic capacity and protects against diet-induced obesity in male mice Scientific Reports, 2021-08-12;11(1):16409. 2021-08-12 [PMID: 34385484] (Bioassay, Human)

Bioinformatics

• Uniprot:
  • Q6P988()