Recombinant Human Nidogen-1/Entactin Protein, CF Summary
Details of Functionality
Measured by the ability of the immobilized protein to support the adhesion of SVEC4‑10 mouse vascular endothelial cells. When 4 x 104 cells/well are added to rhNidogen-1 coated plates (30 µg/mL with 100 µL/well), approximately 40-75% will adhere after one hour at 37 °C. Optimal dilutions should be determined by each laboratory for each application.
Source
Mouse myeloma cell line, NS0-derived human Nidogen-1/Entactin protein Leu29-Lys1114 (Gln1113Arg), with an N-terminal 9-His tag
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.
Applications/Dilutions
Dilutions
Bioactivity
Theoretical MW
120 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
130 kDa, reducing conditions
Publications
Read Publications using 2570-ND in the following applications:
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 under reducing conditions and visualized by silver stain
Reconstitution Instructions
Reconstitute at 100 μg/mL in sterile PBS.
Notes
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Human Nidogen-1/Entactin Protein, CF
enactin
Entactin
Entactin-1
NID1
NID-1
NIDentactin
nidogen (enactin)
nidogen 1
Nidogen1
Nidogen-1
Background
Nidogen-1 (also entactin) is a 150 kDa, secreted, monomeric glycoprotein that serves as a major linking component of basement membranes (1 - 4). It is synthesized as a 1247 amino acid (aa) precursor with a 28 aa signal sequence and a 1219 aa mature protein. The molecule is modular in structure with five distinct regions. There are three globular domains (G1-3) separated by a mucin region and an extended rod-shaped segment (5 - 7). The N-terminal globular domain (G1) is 200 aa in length and seemingly unrelated to any known motif (8). The mucin region is nearly 160 aa in length and presumably O-glycosylated (2, 8). G2 and G3 are both approximately 300 aa in length. G2 is described as a Nidogen ( beta -barrel) domain, while C-terminal G3 assumes a beta -propeller configuration (1). The 250 aa rod-shaped segment has multiple EGF-like motifs and two thyroglobulin type 1 domains. Functionally, G1 is reported to bind type IV collagen (2, 7). The mucin region contains a short peptide that ligates alpha 3 beta 1 integrins (9, 10). G2 interacts with perlecan, and an RGD motif in the rod-shaped segment serves as a binding site for alpha v beta 3 integrins (9, 10). Finally, G3 is associated with laminin binding (2, 7). As a full-length molecule, the multiple extracellular matrix-binding sites of Nidogen-1 are well positioned to serve as anchor sites for basement membrane molecules. Nidogen-1 also undergoes proteolytic processing by at least two MMPs, MMP7 and MMP19 (10, 11). While this destroys the integrity of Nidogen-associated matrices, it also generates peptide fragments that are capable of inducing neutrophil chemotaxis and phagocytosis (10). Nidogen-2 is related to Nidogen-1 (≈ 50% aa identity) and shares many of the same adhesive properties as Nidogen-1 (12). Both bind perlecan plus collagens I and IV. Nidogen-2, however, does not bind fibulin-1 or 2, and shows only modest interaction with laminin. Thus, although coexpressed, Nidogen-2 serves as only a partial substitute for Nidogen-1 (2, 12). Human Nidogen-1 is 85% aa identical to both mouse and rat Nidogen-1, and 88% aa identical to canine Nidogen-1.
Hohenester, E. and J. Engel (2002) Matrix Biol. 21:115.
Miosge, N. et al. (2001) Histochem. J. 33:523.
Charonis, A. et al. (2005) Curr. Med. Chem. 12:1495.
Timpl, R. and J.C. Brown (1996) BioEssays 18:123.
Nagayoshi, T. et al. (1989) DNA 8:581.
Zimmerman, K. et al. (1995) Genomics 27:245.
Fox, J.W. et al. (1991) EMBO J. 10:3137.
Mayer, U. et al. (1995) Eur. J. Biochem. 227:681.
Gresham, H.D. et al. (1996) J. Biol. Chem. 271:30587.
Dong, L-J. et al. (1995) J. Biol. Chem. 270:15383.
Titz, B. et al. (2004) Cell. Mol. Life Sci. 61:1826.
The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.
=
÷
Review this Product
Be the first to review our Recombinant Human Nidogen-1/Entactin Protein, CF and receive a gift card or discount.