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Human Nidogen-1/Entactin Antibody

Catalog #: AF2570
18 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF2570
AF2570-SP
Nidogen‑1/Entactin in Human Heart.
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Citations (18)
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Human Nidogen-1/Entactin Antibody Summary

Species Reactivity
Human
Specificity
Detects human Nidogen‑1/Entactin in direct ELISAs and Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Nidogen‑1/Entactin
Leu29-Lys1114 (Gln1113Arg)
Accession # AAH45606
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Human Nidogen‑1/Entactin (Catalog # 2570-ND)
Immunohistochemistry
5-15 µg/mL
See below

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.

Scientific Data

Immunohistochemistry Nidogen‑1/Entactin antibody in Human Heart by Immunohistochemistry (IHC-P). View Larger

Nidogen‑1/Entactin in Human Heart. Nidogen‑1/Entactin was detected in immersion fixed paraffin-embedded sections of human heart using Goat Anti-Human Nidogen‑1/Entactin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2570) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the sarcolemma of cardiomyocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Western Blot Detection of Human Nidogen-1/Entactin by Western Blot View Larger

Detection of Human Nidogen-1/Entactin by Western Blot Degradation of nidogen-1 by PSA.PSA cleaved both mouse nidogen-1 in Matrigel and human recombinant nidogen-1. Mass spectrometry analysis identified nidogen-1 in the silver stained gel bands (arrows, left panel). Nidogen-1 bands of 140 kDa (A) and 110 kDa (B) disappeared and fragments of 90 kDa (C) and 55 kDa (D) appeared after 48 h incubation of diluted Matrigel with PSA at 37°C. Nidogen-1 cleavage by PSA in Matrigel was visualized by Western blotting with anti-nidogen-1 polyclonal antibody (middle panel). PSA (1 µM) cleaved recombinant human nidogen-1 (0.5 µM) into two fragments 85 kDa (arrow with *) and 55 kDa (arrow with **) during 20 h incubation at 37°C (right panel). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0107819), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Human Nidogen-1/Entactin by Western Blot View Larger

Detection of Human Nidogen-1/Entactin by Western Blot Cleavage of protein substrates by PSA.(A) Characterization of the proteolytic activity of PSA (0.2 µM) during 22 h incubation towards different protein substrates (1 µM each, except 0.5 µM semenogelin I) by SDS-PAGE and silver staining. Approximate molecular weights of the proteins are: PSA (28 kDa), semenogelin I (50 kDa), semenogelin II (63 kDa), fibronectin (220 kDa), galectin-3 (26 kDa), IGFBP-3 (30 kDa) and nidogen-1 (130 kDa). The lanes in which ∼50% of the proteins were cleaved are bordered. (B) 1 µM MMP-3 (22 kDa), but not PSA, cleaved 1 µM plasminogen (88 kDa). Also 0.5 M fibronectin was incubated with 1 µM PSA as a control (SDS-PAGE with silver staining). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0107819), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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

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Citations for Human Nidogen-1/Entactin Antibody

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.

18 Citations: Showing 1 - 10
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  1. Nidogen-1 expression is associated with overall survival and temozolomide sensitivity in low-grade glioma patients
    Authors: Baiwei Zhang, Cheng Xu, Junfeng Liu, Jinsheng Yang, Qinglei Gao, Fei Ye
    Aging (Albany NY)
  2. The extracellular matrix complexity of idiopathic epiretinal membranes and the bilaminar arrangement of the associated internal limiting membrane in the posterior retina
    Authors: Altera A, Tosi GM, Regoli M et al.
    Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie
  3. A comprehensive proteomics study on platelet concentrates: Platelet proteome, storage time and Mirasol pathogen reduction technology
    Authors: V Salunkhe, IM De Cuyper, P Papadopoul, PF van der Me, BB Daal, M Villa-Faja, D de Korte, TK van den Be, L Gutiérrez
    Platelets, 2018-03-19;0(0):1-12.
  4. Cardiac progenitor cell-derived extracellular vesicles promote angiogenesis through both associated- and co-isolated proteins
    Authors: Roefs MT, Bauzá-Martinez J, van de Wakker SI et al.
    Communications biology
  5. Corneal epithelial basement membrane assembly is mediated by epithelial cells in coordination with corneal fibroblasts during wound healing
    Authors: Shiju TM, Sampaio LP, Hilgert GSL, Wilson SE
    Molecular vision
  6. Development of stromal differentiation patterns in heterotypical models of artificial corneas generated by tissue engineering
    Authors: Blanco-Elices C, Morales-�lvarez C, Chato-Astrain J et al.
    Frontiers in Bioengineering and Biotechnology
  7. Expression of Basement Membrane Molecules by Wharton Jelly Stem Cells (WJSC) in Full-Term Human Umbilical Cords, Cell Cultures and Microtissues
    Authors: Sánchez-Porras D, Durand-Herrera D, Carmona R et al.
    Cells
  8. BET protein inhibitor JQ1 downregulates chromatin accessibility and suppresses metastasis of gastric cancer via inactivating RUNX2/NID1 signaling
    Authors: S Zhou, S Zhang, L Wang, S Huang, Y Yuan, J Yang, H Wang, X Li, P Wang, L Zhou, J Yang, Y Xu, H Gao, Y Zhang, Y Lv, X Zou
    Oncogenesis, 2020-03-10;9(3):33.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  9. Immune-mediated ECM depletion improves tumour perfusion and payload delivery
    Authors: YL Yeow, VR Kotamraju, X Wang, M Chopra, N Azme, J Wu, TD Schoep, DS Delaney, K Feindel, J Li, KM Kennedy, WM Allen, BF Kennedy, I Larma, DD Sampson, LM Mahakian, BZ Fite, H Zhang, T Friman, AP Mann, FA Aziz, MP Kumarasing, M Johansson, HC Ee, G Yeoh, L Mou, KW Ferrara, H Billiran, R Ganss, E Ruoslahti, J Hamzah
    EMBO Mol Med, 2019-11-11;0(0):e10923.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  10. Descemet's Membrane Modulation of Posterior Corneal Fibrosis
    Authors: CS Medeiros, P Saikia, RC de Oliveir, L Lassance, MR Santhiago, SE Wilson
    Invest. Ophthalmol. Vis. Sci., 2019-03-01;60(4):1010-1020.
    Species: Rabbit
    Sample Types: Whole Tissue
    Applications: IHC-Fr
  11. IL-1 and TGF-? Modulation of Epithelial Basement Membrane Components Perlecan and Nidogen Production by Corneal Stromal Cells
    Authors: P Saikia, S Thangavadi, CS Medeiros, L Lassance, RC de Oliveir, SE Wilson
    Invest. Ophthalmol. Vis. Sci., 2018-11-01;59(13):5589-5598.
    Species: Rabbit
    Sample Types: Cell Extracts
    Applications: Western Blot
  12. Posterior stromal cell apoptosis triggered by mechanical endothelial injury and basement membrane component nidogen-1 production in the cornea
    Authors: CS Medeiros, L Lassance, P Saikia, M Santhiago, SE Wilson
    Exp. Eye Res., 2018-03-27;0(0):.
    Species: Rabbit
    Sample Types: Whole Cells
    Applications: IHC
  13. Proteolytic activity of prostate-specific antigen (PSA) towards protein substrates and effect of peptides stimulating PSA activity.
    Authors: Mattsson J, Ravela S, Hekim C, Jonsson M, Malm J, Narvanen A, Stenman U, Koistinen H
    PLoS ONE, 2014-09-19;9(9):e107819.
    Species: Human
    Sample Types: Protein
    Applications: Western Blot
  14. Hematopoietic stem cell cytokines and fibroblast growth factor-2 stimulate human endothelial cell-pericyte tube co-assembly in 3D fibrin matrices under serum-free defined conditions.
    Authors: Smith A, Bowers S, Stratman A, Davis G
    PLoS ONE, 2013-12-31;8(12):e85147.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  15. Development and characterization of a high-throughput in vitro cord formation model insensitive to VEGF inhibition.
    Authors: Falcon B, O'Clair B, McClure D, Evans G, Stewart J, Swearingen M, Chen Y, Allard K, Lee L, Neote K, McEwen D, Uhlik M, Chintharlapalli S
    J Hematol Oncol, 2013-04-27;6(0):31.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  16. Pericyte recruitment during vasculogenic tube assembly stimulates endothelial basement membrane matrix formation.
    Authors: Stratman AN, Malotte KM, Mahan RD, Davis MJ, Davis GE
    Blood, 2009-10-12;114(24):5091-101.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: ICC, Western Blot
  17. TGF beta 1 and TGF beta 2 proteins in corneas with and without stromal fibrosis: Delayed regeneration of apical epithelial growth factor barrier and the epithelial basement membrane in corneas with stromal fibrosis
    Authors: Carlos de Oliveira R, Tye G, Sampaio L et al.
    Experimental Eye Research
  18. Epithelial Basement Membrane Regeneration After PRK-Induced Epithelial-Stromal Injury in Rabbits: Fibrotic Versus Non-fibrotic Corneal Healing
    Authors: de Oliveira RC, Sampaio LP, Shiju TM et al.
    Journal of refractive surgery (Thorofare, N.J. : 1995)

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