Mouse Netrin-1 Antibody

Catalog # Availability Size / Price Qty
AF1109
AF1109-SP

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Detection of Human Netrin-1 by Western Blot
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Product Details
Citations (13)
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Mouse Netrin-1 Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse Netrin-1 in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant chicken (rch) Netrin‑1 and recombinant human Netrin-1 is observed and less than 1% cross-reactivity with recombinant mouse (rm) Netrin-G1a and rmNetrin-4 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant mouse Netrin-1
Val22-Ala603
Accession # AAC52971
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.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Mouse Netrin‑1 (Catalog # 1109-N1)
Blockade of Receptor-ligand Interaction
In a functional ELISA, 1-3 µg/mL of this antibody will block 50% of the binding of 100 ng/mL of Recombinant Mouse Netrin-1 (Catalog # 1109-N1) to immobilized Recombinant Rat UNC5H2 Fc Chimera (Catalog # 1006-UN) coated at 5 µg/mL (100 µL/well). At 40 μg/mL, this antibody will block >90% of the binding.
 

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

Western Blot Detection of Human Netrin-1 by Western Blot View Larger

Detection of Human Netrin-1 by Western Blot UNC5A and UNC5C induce caspase-3 activation through DAPK1/PR65b during UPR. (A) Quantification of netrin-1–receptor expression by qRT-PCR (mean + SEM, n = 3). (B–F) Identification of the implicated netrin-1 receptors. HepaRG cells were transfected with siRNAs and treated with DTT or not for 4 hours (mock). (B) Assessment of XBP1 mRNA splicing by RT-PCR. Representative result, n = 3. (C) Assessment of netrin-1 protein knockdown by immunoblotting. Representative result, n = 3. (D and E) Assessment of transcript knockdown efficiencies. Graphs indicate (D) UNC5A and (E) UNC5C mRNA levels in siRNA-treated cells in comparison with control siRNA-treated cells (mean + SEM; n = 3; Mann–Whitney test; P <.05). (F) Caspase-3 activation is reversed by UNC5A or UNC5C knockdown after UPR induction. Graph indicates caspase-3 activity ratio of DTT vs untreated cells for each condition (mean + SEM; n = 3; Mann–Whitney test; P <.05). (G–J) Identification of the downstream signaling pathway. HepaRG cells were transfected with siRNAs and treated with DTT or not for 4 hours (mock). (G) Assessment of XBP1 mRNA splicing by RT-PCR. Representative result, n = 3. (H) Evaluation of netrin-1, DAPK1, and PR65 beta depletion by immunoblotting. Representative result, n = 3. (I) Caspase-3 activation is reversed by DAPK1 or PR65 beta knockdown. Graph indicates the caspase-3 activity ratio for each condition (mean + SEM; n = 3; Mann–Whitney test; P <.05). (J) PP2A activity is increased by netrin-1 depletion and reversed by reduced expression of PR65 beta. Graph indicates PP2A activity ratio for each condition (mean + SEM; n = 3; Mann–Whitney test; P <.05). *,**, or *** refer to statistical analyses. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28174720), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Human Netrin-1 by Western Blot View Larger

Detection of Human Netrin-1 by Western Blot Netrin-1 protects against cell death during UPR. (A–H) Netrin-1 depletion. (A–E) HepaRG cells were transfected with netrin-1 siRNA, treated with DTT, and harvested in a time course assay. (A) Netrin-1 protein knockdown by siRNA was validated by immunoblot (representative result, n = 3). (B) XBP1 splicing was confirmed by RT-PCR (representative result, n = 3). (C) UPR increases the dependence of cells toward netrin-1 for survival. Graphs indicate the difference in cell death (in percentages) (mean + SEM; n = 3; Mann–Whitney test; P <.05). (D) UPR increases the sensitivity of cells toward netrin-1 for caspase-3 activation. Graphs indicate caspase-3 activity ratios (mean + SEM; n = 3; Mann–Whitney test; P <.05). (E) Netrin-1 depletion increases apoptosis. Graph indicates apoptotic cell death ratio by propidium iodide staining and flow cytometry (mean + SEM; n = 3; Mann–Whitney test; P <.05). (F–H) Netrin-1 inhibition using a neutralizing antibody. HepaRG cells were seeded and treated with a control (H4) or anti–netrin-1 antibody (2F5) the same day. Cells were treated with DTT 3 days after addition and harvested after the indicated time points. (F) Assessment of XBP1 mRNA splicing (representative result, n = 3). (G) Netrin-1 neutralization enhances caspase-3 activity. Graph indicates the caspase-3 activity ratios between 2F5 and H4-treated cells (mean + SEM; n = 3; Mann–Whitney test; P <.05). (H) Netrin-1 depletion increases apoptosis. Graph indicates the apoptotic cell death ratio as assessed after propidium iodide staining and flow cytometry of netrin-1–depleted and control cells (mean + SEM; n = 3; Mann–Whitney test; P <.05). (I–M) Netrin-1 forced expression. HepaRG cells were transfected with control (VR1-HA) and netrin-1 (netrin-1–HA) vectors, treated with DTT, and harvested. (I) Netrin-1 expression was assessed by immunoblotting. Representative result, n = 3. (J) XBP1 mRNA splicing was quantified by RT-PCR at the indicated time points. Representative result, n = 3. (K) UPR increases the sensitivity of cells toward netrin-1 for protection against cell death in time course assays. Graph indicates the difference in cell death (in percentages) between netrin-1–overexpressing cells and control cells (mean + SEM; n = 3; Mann–Whitney test; P <.05). (L) UPR increases the sensitivity of cells toward netrin-1 for caspase-3 activation in time course assays. Graph indicates the ratios of caspase-3 activation levels among netrin-1–overexpressing and control cells (mean + SEM; n = 3; Mann–Whitney test; P <.05). (M) Netrin-1 overexpression decreases apoptosis in a time course assay. Graph indicates the apoptotic cell death ratio as assessed after propidium iodide staining and flow cytometry of netrin-1–overexpressing and control cells (mean + SEM; n = 3; Mann–Whitney test; P <.05). *,**, or *** refer to statistical analyses. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28174720), licensed under a CC-BY license. Not internally tested by R&D Systems.

Western Blot Detection of Mouse Netrin-1 by Western Blot View Larger

Detection of Mouse Netrin-1 by Western Blot Netrin-1 reverts UPR-induced caspase-3 activation in netrin-1 transgenic mice. Netrin-1 (FLAG-tagged) transgenic mice or control littermates were treated with Tamoxifen (Tamox), injected with PBS, or 1 mg/kg Tu and killed 24 hours after treatment. (A) Evaluation of netrin-1 protein overexpression by anti-FLAG immunoblotting. (B) Evaluation of netrin-1 protein overexpression by immunohistochemistry. Representative result, n = 5 (wt/tamox/PBS), n = 6 (wt/tamox/Tu), n = 3 (netrin-1/tamox/PBS), n = 5 (netrin-1/tamox/Tu for the whole figure). Background level was assessed using an isotype control antibody. Magnification: ×20. (C) Liver pictures 24 hours after treatment. (D) Evaluation of eIF2 alpha phosphorylation by immunoblotting. (E) Caspase-3 activation after Tu treatment is reversed in netrin-1–expressing transgenic mice. Graph indicates fold changes in caspase-3 activity compared with control mice (mean + SEM; Mann–Whitney test; P <.05). (F) The number of apoptotic cells is decreased in netrin-1 transgenic mice after Tu treatment. Graph indicates TUNEL positive cells/mm2 for each group (mean + SEM; Mann–Whitney test; P <.05). See also Supplementary Figure 6. *,**, or *** refer to statistical analyses. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28174720), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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: Netrin-1

Mouse Netrin-1 is a member of the laminin-related family of axon-guidance molecules, collectively referred to as Netrins (netr is Sanskrit for "one who guides"). The molecule's cDNA encodes a 603 amino acid (aa) protein precursor that has structural similarity to the N-terminal gamma -chain of laminin. It contains a globular domain, three EGF repeats, and a C-terminal heparin-binding domain. Mouse Netrin-1 shares 52% aa identity with mouse Netrin-3, and 98% and 87% aa identity with human and chicken Netrin-1, respectively. Cells reported to express Netrin-1 in the embryo include cells of the floor plate, ventricular zone of the spinal cord, the brain, the ganglionic eminence, and parts of the diencephalon. Netrins were first identified for promoting the outgrowth of commissural axons and are also involved in helping migrating cells and axonal growth cones navigate to their targets. Netrins can provide both attractive and repulsive cues to neurons, depending on the receptors present and cellular context. In the adult, Netrin-1 is likely involved in axon regeneration in peripheral nerves. Netrin-1 has also been shown to be expressed outside of the nervous system and to be involved in development of such tissues as the pancreas, lung, bowel, bone and mammary gland. In non-neural organogenesis, Netrin‑1 provides an adhesive rather than guidance function. The DCC (deleted in colorectal carcinoma), Neogenin, the UNC5 family of receptors, and the adenosine A2b receptors are proposed to be functional receptors for Netrin-1 (1-7).

References
  1. Puschel, A. (1999) Mech. Dev. 83:65.
  2. Hedgecock, E. and C. Norris (1997) Trends Genet. 13:251.
  3. Kappler, J. et al. (2000) Biochem. Biophys. Res. Commun. 271:287.
  4. Madison, R. et al. (2000) Exp. Neurology 161:563.
  5. Srinivasan, K. et al. (2003) Dev. Cell 4:371.
  6. Livesey, F.J. (1999) Cell Mol. Life Sci. 56:62.
  7. Corset, V. et al. (2000) Nature 407:747.
Entrez Gene IDs
9423 (Human); 18208 (Mouse); 396389 (Chicken)
Alternate Names
netrin 1; Netrin1; Netrin-1; NTN1; NTN1Lnetrin 1, mouse, homolog of

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Product Specific Notices

This product or the use of this product is covered by U.S. Patents owned by The Regents of the University of California. This product is for research use only and is not to be used for commercial purposes. Use of this product to produce products for sale or for diagnostic, therapeutic or drug discovery purposes is prohibited. In order to obtain a license to use this product for such purposes, contact The Regents of the University of California.

U.S. Patent # 5,565,331, 6,096,866, 6,017,714, 6,309,638, 6,670,451, and other U.S. and international patents pending.

Citations for Mouse Netrin-1 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.

13 Citations: Showing 1 - 10
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  1. Netrin1 patterns the dorsal spinal cord through modulation of Bmp signaling
    Authors: Alvarez, S;Gupta, S;Honeychurch, K;Mercado-Ayon, Y;Kawaguchi, R;Butler, SJ;
    bioRxiv : the preprint server for biology
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  2. Netrin-1 promotes the immunosuppressive activity of MDSCs in colorectal cancer
    Authors: X Xia, Z Mao, W Wang, J Ma, J Tian, S Wang, K Yin
    Cancer Immunology Research, 2023-05-03;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  3. Restoring bone marrow niche function rejuvenates aged hematopoietic stem cells by reactivating the DNA Damage Response
    Authors: P Ramalingam, MC Gutkin, MG Poulos, T Tillery, C Doughty, A Winiarski, AG Freire, S Rafii, D Redmond, JM Butler
    Nature Communications, 2023-04-10;14(1):2018.
    Species: Mouse
    Sample Types: Whole Cells, Whole Tissue
    Applications: ICC, IHC
  4. Allosteric potentiation of GABAA receptor single-channel conductance by netrin-1 during neuronal-excitation-induced inhibitory synaptic homeostasis
    Authors: ES Chan, Y Ge, YW So, YF Bai, L Liu, YT Wang
    Cell Reports, 2022-11-01;41(5):111584.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  5. Endothelial Unc5B controls blood-brain barrier integrity
    Authors: K Boyé, LH Geraldo, J Furtado, L Pibouin-Fr, M Poulet, D Kim, B Nelson, Y Xu, L Jacob, N Maissa, D Agalliu, L Claesson-W, SL Ackerman, A Eichmann
    Nature Communications, 2022-03-04;13(1):1169.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  6. The expression of Netrin-1 in the MIA-induced osteoarthritic temporomandibular joint in mice
    Authors: M Xiao, Z Hu, H Jiang, C Li, H Guo, W Fang, X Long
    Scientific Reports, 2021-08-03;11(1):15695.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: IHC
  7. Netrin 1 mediates protective effects exerted by insulin-like growth factor 1 on cochlear hair cells
    Authors: K Yamahara, T Nakagawa, J Ito, K Kinoshita, K Omori, N Yamamoto
    Neuropharmacology, 2017-04-01;119(0):26-39.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Neutralization
  8. Netrin-1 Protects Hepatocytes Against Cell Death Through Sustained Translation During the Unfolded Protein Response
    Authors: T Lahlali, ML Plissonnie, C Romero-Lóp, M Michelet, B Ducarouge, A Berzal-Her, F Zoulim, P Mehlen, R Parent
    Cell Mol Gastroenterol Hepatol, 2016-01-09;2(3):281-301.e9.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  9. Netrin-1 promotes adipose tissue macrophage retention and insulin resistance in obesity.
    Authors: Ramkhelawon B, Hennessy E, Menager M, Ray T, Sheedy F, Hutchison S, Wanschel A, Oldebeken S, Geoffrion M, Spiro W, Miller G, McPherson R, Rayner K, Moore K
    Nat Med, 2014-03-02;20(4):377-84.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-P
  10. Endothelial expression of guidance cues in vessel wall homeostasis dysregulation under proatherosclerotic conditions.
    Authors: van Gils, Janine M, Ramkhelawon, Bhama, Fernandes, Luciana, Stewart, Merran C, Guo, Liang, Seibert, Tara, Menezes, Gustavo, Cara, Denise C, Chow, Camille, Kinane, T Bernar, Fisher, Edward A, Balcells, Mercedes, Alvarez-Leite, Jacqueli, Lacy-Hulbert, Adam, Moore, Kathryn
    Arterioscler Thromb Vasc Biol, 2013-02-21;33(5):911-9.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: ICC, Western Blot
  11. Netrin-1 promotes glioblastoma cell invasiveness and angiogenesis by multiple pathways including activation of RhoA, cathepsin B, and cAMP-response element-binding protein.
    Authors: Shimizu A, Nakayama H, Wang P, Konig C, Akino T, Sandlund J, Coma S, Italiano J, Mammoto A, Bielenberg D, Klagsbrun M
    J Biol Chem, 2012-11-29;288(4):2210-22.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  12. Interfering with multimerization of netrin-1 receptors triggers tumor cell death.
    Authors: Mille F, Llambi F, Guix C, Delloye-Bourgeois C, Guenebeaud C, Castro-Obregon S, Bredesen DE, Thibert C, Mehlen P
    Cell Death Differ., 2009-06-19;16(10):1344-51.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: ICC
  13. Distinct roles of Rac1/Cdc42 and Rho/Rock for axon outgrowth and nucleokinesis of precerebellar neurons toward netrin 1.
    Authors: Causeret F, Hidalgo-Sanchez M, Fort P, Backer S, Popoff MR, Gauthier-Rouviere C, Bloch-Gallego E
    Development, 2004-05-19;131(12):2841-52.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot

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