Recombinant Human Nodal Protein, CF

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

Recombinant Human Nodal Protein, CF Summary

• Details of Functionality: Measured by its ability to induce Smad2 phosphorylation in P19 mouse embryonal carcinoma cells. Yeo, C. et al. (2001) Mol. Cell. 7:949. Approximately 100 ng/mL of Recombinant Human Nodal can effectively induce Smad2 phosphorylation.
• Source: E. coli-derived human Nodal protein
  His238-Leu347, with an N-terminal Met
• Accession #: Q96S42
• N-terminal Sequence: Met
• Structure / Form: Disulfide-linked homodimer
• Protein/Peptide Type: Recombinant Proteins
• Gene: NODAL
• Purity: >97%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 12.9 kDa (monomer).
  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.

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 Acetonitrile and TFA.
• Purity: >97%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Reconstitution Instructions: Reconstitute at 100 μg/mL in sterile 4 mM HCl.

Notes

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

Alternate Names for Recombinant Human Nodal Protein, CF

• BMP-16
• MGC138230
• nodal homolog (mouse)
• nodal homolog
• Nodal
• nodal, mouse, homolog

Background

Nodal is a 13 kDa member of the TGF-beta superfamily of molecules (1-3). Its name derives from the fact that it is expressed in the primitive node of the developing embryo. In human, it is synthesized as a 347 amino acid (aa) preproprecursor that contains a 26 aa signal sequence, a 211 aa prodomain, and a 110 aa mature region (4, 5). Consistent with its TGF-beta superfamily membership, it exists as a disulfide-linked homodimer and would be expected to demonstrate a cysteine-knot motif (1, 5, 6). Mature human Nodal is 99%, 98%, 96% and 98% aa identical to mature canine, rat, bovine and mouse Nodal, respectively. Nodal signals through two receptor complexes, both of which contain members of the TGF-beta family of Ser/Thr kinase receptors. The first receptor complex is composed of ActRIB/ALK4 and ActRIIB. Nodal interaction with the ligand-binding subunit ALK4 requires the participation of fucosylated Cripto, a member of the EGF-CFC gene family. The second complex is composed of ALK7 and ActRIIB, and does not require Cripto interaction (7-9). Nodal signaling is regulated by a number of inhibitors. Nodal is reported to form nonsignaling disulfide-linked heterodimers with BMP-3, -4 and -7 (2, 6). There are also at least three secreted factors that block Nodal activation of its receptor. The first is Cerberus, a member of the DAN family of proteins. The last two are TGF-beta family members named Lefty-A and B (2, 11). The exact mechanisms employed are unclear, but Leftys are apparently induced by nodal and can work at a considerable distance from their origin (2). Nodal is known to induce both mesoderm and endoderm, participate in anterior-posterior positioning, and through ALK7, induce apoptosis in susceptible cell populations (2, 8, 12).

1. Kishigami, S. and Y. Mishina (2005) Cytokine Growth Factor Rev. 16:265.
2. Schier, A.F. (2003) Annu. Rev. Cell Dev. Biol. 19:589.
3. Whitman, M. and M. Mercola (2001) Sci. STKE. Jan 9;(64):RE1.
4. Gebbia, M. et al. (1997) Nat. Genet. 17:305.
5. Hart, A.H. et al. (2005) Biochem. Biophys. Res. Commun. 333:1361.
6. Yeo, C-Y. and M. Whitman (2001) Mol. Cell 7:949.
7. Yan, Y-T. et al. (2002) Mol. Cell. Biol. 22:4439.
8. Munir, S. et al. (2004) J. Biol. Chem. 279:31277.
9. Shen, M.M. and A.F. Schier (2000) Trends Genet. 16:303.
10. Lah, M. et al. (1999) Genomics 55:364.
11. Kosaki, K. et al. (1999) Am. J. Hum. Genet. 64:712.
12. Zhou, X. et al. (1993) Nature 361:543.

Publications for Nodal (3218-ND/CF)(15)

Publications using 3218-ND/CF

• K Harpelunde, JE Nielsen, B Grønkær To, UN Joensen, LJ Rasmussen, M Blomberg J, RT Mitchell, A Juul, E Rajpert-De, A Jørgensen Influence of Nodal signalling on pluripotency factor expression, tumour cell proliferation and cisplatin-sensitivity in testicular germ cell tumours BMC Cancer, 2020;20(1):349. 2020 [PMID: 32326899] (Bioassay, Human)
• SC Vermilyea, A Babinski, N Tran, S To, S Guthrie, JH Kluss, JK Schmidt, GJ Wiepz, MG Meyer, ME Murphy, MR Cookson, ME Emborg, TG Golos In Vitro CRISPR/Cas9-Directed Gene Editing to Model LRRK2 G2019S Parkinson&#039;s Disease in Common Marmosets Sci Rep, 2020;10(1):3447. 2020 [PMID: 32103062] (Cell Culture, Simian)
• P Ray, AS Chin, KE Worley, J Fan, G Kaur, M Wu, LQ Wan Intrinsic cellular chirality regulates left-right symmetry breaking during cardiac looping Proc. Natl. Acad. Sci. U.S.A., 2018;0(0):. 2018 [PMID: 30459275] (Bioassay, Chicken)
• A Jørgensen, J Macdonald, JE Nielsen, KR Kilcoyne, S Perlman, L Lundvall, L Langhoff T, K Juul Hare, H Frederikse, AM Andersson, NE Skakkebæk, A Juul, RM Sharpe, E Rajpert-De, RT Mitchell Nodal Signaling Regulates Germ Cell Development and Establishment of Seminiferous Cords in the Human Fetal Testis Cell Rep, 2018;25(7):1924-1937.e4. 2018 [PMID: 30428358] (Bioassay, Human)
• Y Jiang, H Li, Y Wang, T Tian, Y He, Y Jin, C Han, X Jin, F Zhang, E Morii ALDH enzyme activity is regulated by Nodal and histamine in the A549 cell line Oncol Lett, 2017;14(6):6955-6961. 2017 [PMID: 29181106] (Bioassay, Human)
• CR Ball, F Oppel, KR Ehrenberg, TD Dubash, SM Dieter, CM Hoffmann, U Abel, F Herbst, M Koch, J Werner, F Bergmann, N Ishaque, M Schmidt, C von Kalle, C Scholl, S Fröhling, B Brors, W Weichert, J Weitz, H Glimm Succession of transiently active tumor-initiating cell clones in human pancreatic cancer xenografts EMBO Mol Med, 2017;0(0):. 2017 [PMID: 28526679] (Bioassay, Human)
• Taroh Kinoshita A GPI processing phospholipase A2, PGAP6, modulates Nodal signaling in embryos by shedding CRIPTO J. Cell Biol., 2016;215(5):705-718. 2016 [PMID: 27881714] (Bioassay, Human)
• Fuerer C, Nostro M, Constam D Nodal.Gdf1 heterodimers with bound prodomains enable serum-independent nodal signaling and endoderm differentiation. J Biol Chem, 2014;289(25):17854-71. 2014 [PMID: 24798330] (Western Blot, Human)
• Esquivies L, Blackler A, Peran M, Rodriguez-Esteban C, Izpisua Belmonte J, Booker E, Gray P, Ahn C, Kwiatkowski W, Choe S Designer nodal/BMP2 chimeras mimic nodal signaling, promote chondrogenesis, and reveal a BMP2-like structure. J Biol Chem, 2014;289(3):1788-97. 2014 [PMID: 24311780] (Bioassay, Human)
• Quail , D F, Zhang , G, Findlay , S D, Hess , D A, Postovit , L-M Nodal promotes invasive phenotypes via a mitogen-activated protein kinase-dependent pathway. Oncogene, 2014;33(4):461-73. 2014 [PMID: 23334323] (Bioassay, Human)
• Wilson TR, Fridlyand J, Yan Y, Penuel E, Burton L, Chan E, Peng J, Lin E, Wang Y, Sosman J, Ribas A, Li J, Moffat J, Sutherlin DP, Koeppen H, Merchant M, Neve R, Settleman J Widespread potential for growth-factor-driven resistance to anticancer kinase inhibitors. Nature, 2012;487(7408):505-9. 2012 [PMID: 22763448] (Bioassay, Human)
• Castonguay R, Werner ED, Matthews RG, Presman E, Mulivor AW, Solban N, Sako D, Pearsall RS, Underwood KW, Seehra J, Kumar R, Grinberg AV Soluble Endoglin Specifically Binds Bone Morphogenetic Proteins 9 and 10 via Its Orphan Domain, Inhibits Blood Vessel Formation, and Suppresses Tumor Growth. J. Biol. Chem., 2011;286(34):30034-46. 2011 [PMID: 21737454] (Surface Plasmon Resonance, Human)
• Herrera B, Inman GJ, A rapid and sensitive bioassay for the simultaneous measurement of multiple bone morphogenetic proteins. Identification and quantification of BMP4, BMP6 and BMP9 in bovine and human serum. BMC Cell Biol., 2009;10(0):20. 2009 [PMID: 19298647] (Bioassay, Mouse)
• Watanabe K, Hamada S, Bianco C, Mancino M, Nagaoka T, Gonzales M, Bailly V, Strizzi L, Salomon DS Requirement of glycosylphosphatidylinositol anchor of Cripto-1 for trans activity as a Nodal co-receptor. J. Biol. Chem., 2007;282(49):35772-86. 2007 [PMID: 17925387] (Bioassay, Human)
• McLean AB, D&amp;amp;apos;Amour KA, Jones KL, Krishnamoorthy M, Kulik MJ, Reynolds DM, Sheppard AM, Liu H, Xu Y, Baetge EE, Dalton S Activin a efficiently specifies definitive endoderm from human embryonic stem cells only when phosphatidylinositol 3-kinase signaling is suppressed. Stem Cells, 2007;25(1):29-38. 2007 [PMID: 17204604] (Bioassay, Human)

Bioinformatics

• Gene Symbol: NODAL
• Uniprot:
  • Q96S42()