Human Nidogen-1/Entactin Antibody

(2 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Nidogen‑1/Entactin in ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant human Nidogen-2 is observed.
  • Source
    Monoclonal Mouse IgG1 Clone # 302117
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Nidogen‑1/Entactin
    Leu29-Lys1114 (Gln1113Arg)
    Accession # AAH45606.1
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    Recombinant Human Nidogen‑1/Entactin (Catalog # 2570-ND)
  • Immunohistochemistry
    8-25 µg/mL
    Immersion fixed paraffin-embedded sections of human heart
  • Immunocytochemistry
    8-25 µg/mL
    See below
    • Human Nidogen-1/Entactin Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    2-8 µg/mL 
    Human Nidogen‑1/Entactin Antibody (Catalog # MAB2570)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Human Nidogen-1/Entactin Biotinylated Antibody (Catalog # BAF2570)
  • ELISA Standard
     
    Recombinant Human Nidogen-1/Entactin Protein, CF (Catalog # 2570-ND)
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Immunocytochemistry

Nidogen‑1/Entactin in Human Chondrocytes. Nidogen‑1/Entactin was detected in immersion fixed human mesenchymal stem cells differentiated into chondrocytes using Mouse Anti-Human Nidogen‑1/Entactin Monoclonal Antibody (Catalog # MAB2570) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & 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.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Nidogen-1/Entactin

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, MMP-7 and MMP-19 (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 shares 85% aa sequence identity with both mouse and rat Nidogen-1, and 88% aa sequence identity with canine Nidogen-1.

  • References:
    1. Hohenester, E. and J. Engel (2002) Matrix Biol. 21:115. 
    2. Miosge, N. et al. (2001) Histochem. J. 33:523.
    3. Charonis, A. et al. (2005) Curr. Med. Chem. 12:1495.
    4. Timpl, R. and J.C. Brown (1996) BioEssays 18:123. 
    5. Nagayoshi, T. et al. (1989) DNA 8:581. 
    6. Zimmerman, K. et al. (1995) Genomics 27:245. 
    7. Fox, J.W. et al. (1991) EMBO J. 10:3137.
    8. Mayer, U. et al. (1995) Eur. J. Biochem. 227:681.
    9. Gresham, H.D. et al. (1996) J. Biol. Chem. 271:30587.
    10. Dong, L-J. et al. (1995) J. Biol. Chem. 270:15383.
    11. Titz, B. et al. (2004) Cell. Mol. Life Sci. 61:1826.
    12. Kohfeldt, K. et al. (1998) J. Mol. Biol. 282:99.
  • Entrez Gene IDs:
    4811 (Human)
  • Alternate Names:
    enactin; Entactin; Entactin-1; NID1; NID-1; NIDentactin; nidogen (enactin); nidogen 1; Nidogen1; Nidogen-1
Related Research Areas
Citations:

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.

2 Citations: Showing 1 - 2
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Species
Applications
Sample Type
  1. Enhanced wound healing, kinase and stem cell marker expression in diabetic organ-cultured human corneas upon MMP-10 and cathepsin F gene silencing.
    Authors: Saghizadeh M, Epifantseva I, Hemmati D, Ghiam C, Brunken W, Ljubimov A
    Invest Ophthalmol Vis Sci, 2013;54(13):8172-80.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
  2. Normalization of wound healing and diabetic markers in organ cultured human diabetic corneas by adenoviral delivery of c-Met gene.
    Authors: Saghizadeh M, Kramerov AA, Yu FS, Castro MG, Ljubimov AV
    Invest. Ophthalmol. Vis. Sci., 2010;51(4):1970-80.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
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