Name: Human Netrin-4 Antibody
Brand: R&D Systems
SKU: AF1254
Price: 459 USD

Species Reactivity

Human

Specificity

Detects human Netrin-4 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 50% cross-reactivity with recombinant mouse Netrin-4 is observed, 15% cross-reactivity with recombinant chicken (rch) Netrin-1 is observed and less than 5% cross‑reactivity with recombinant mouse (rm) Netrin-G1a, Netrin-G2a, and rchNetrin-2 is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Netrin-4
Val26-Lys628
Accession # Q9HB63

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 Netrin‑4 (Catalog # 1254-N4)

Immunohistochemistry

5-15 µg/mL

Immersion fixed paraffin-embedded sections of human kidney

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

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Detection of Human Netrin-4 by Immunohistochemistry Immunohistochemistry of NTN4 expression within NB. Representative light microscopy images of a neuroblastoma sample from a female patient (age > 18M); tumor with retroperotineal location and in disseminated stage. (a) Hematoxylin and eosin staining reveals presence of ganglionar-differentiated cells (yellow arrowhead) and primitive neuroblasts (yellow asterisk). (b) Immunohistochemistry of NTN4 with its corresponding negative control (inset). (c) PCNA staining. (d) NTN4 is expressed preferentially in the endothelium (yellow arrows), as confirmed by CD31 staining (inset). (e) Detail of ganglionar-differentiated cells with strong NTN4 cytoplasmic expression in a characteristic punctuated pattern (yellow arrowhead). (f) Close up image of (b) as indicated, highlighting NTN4 expression within the stroma (yellow asterisk). A, B, C; Scale bar = 40 μm. D, E, F; Scale bar = 10 μm; inset = 10 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30160193), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunohistochemistry

View Larger

Detection of Human Netrin-4 by Immunohistochemistry Immunohistochemistry of NTN4 expression within NB. Representative light microscopy images of a neuroblastoma sample from a female patient (age > 18M); tumor with retroperotineal location and in disseminated stage. (a) Hematoxylin and eosin staining reveals presence of ganglionar-differentiated cells (yellow arrowhead) and primitive neuroblasts (yellow asterisk). (b) Immunohistochemistry of NTN4 with its corresponding negative control (inset). (c) PCNA staining. (d) NTN4 is expressed preferentially in the endothelium (yellow arrows), as confirmed by CD31 staining (inset). (e) Detail of ganglionar-differentiated cells with strong NTN4 cytoplasmic expression in a characteristic punctuated pattern (yellow arrowhead). (f) Close up image of (b) as indicated, highlighting NTN4 expression within the stroma (yellow asterisk). A, B, C; Scale bar = 40 μm. D, E, F; Scale bar = 10 μm; inset = 10 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30160193), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunohistochemistry

View Larger

Detection of Human Netrin-4 by Immunohistochemistry Immunohistochemistry of NTN4 expression within NB. Representative light microscopy images of a neuroblastoma sample from a female patient (age > 18M); tumor with retroperotineal location and in disseminated stage. (a) Hematoxylin and eosin staining reveals presence of ganglionar-differentiated cells (yellow arrowhead) and primitive neuroblasts (yellow asterisk). (b) Immunohistochemistry of NTN4 with its corresponding negative control (inset). (c) PCNA staining. (d) NTN4 is expressed preferentially in the endothelium (yellow arrows), as confirmed by CD31 staining (inset). (e) Detail of ganglionar-differentiated cells with strong NTN4 cytoplasmic expression in a characteristic punctuated pattern (yellow arrowhead). (f) Close up image of (b) as indicated, highlighting NTN4 expression within the stroma (yellow asterisk). A, B, C; Scale bar = 40 μm. D, E, F; Scale bar = 10 μm; inset = 10 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30160193), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

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: Netrin-4

Netrins/UNC-6 (netr: Sanskrit for “one who guides”) are a family of laminin-related small proteins that are involved in neurite outgrowth and axon guidance. Netrins bind to the DCC and UNC5 family of receptors to attract or repel axons. Human Netrin-4 is synthesized as a 628 amino acid (aa) precursor that contains a 19 aa signal sequence, a 443 aa laminin-related region containing an N-terminal laminin globular domain (domain VI) followed by 3 laminin EGF-like repeats, and a 166 aa C‑terminal domain rich in basic aa residues that serves as a heparin binding site. Unlike human Netrin-1 which is reminiscent of the laminin gamma -chain, human Netrin-4 resembles the laminin beta -chain. Netrin-4 has been reported to exist as both a monomer and a dimer. The dimeric form was reported to be the less active of the two. Human Netrin-4 shares 31%, 29% and 25% aa sequence identity with human Netrin-1, 2 and G2, respectively; it shares 89% aa identity with mouse Netrin-4. Netrin-4 has widespread expression, occurring in nervous tissues such as embryonic floor plate and postnatal neurons such as cerebellar granule cells and hippocampal pyramidal cells. It also is found in non-neural tissues, such as adult Bowman’s capsule and medullary tubular epithelium in kidney, and embryonic pancreatic and intestinal epithelium, plus cells of the ureteric bud. Netrin-4 expression is often associated with basement membrane. Netrin-4 has been shown to initiate axon outgrowth from olfactory bulb explants (1‑7).

References

McFarlane, S. (2000) Biochem. Cell. Biol. 78:563.
Kennedy, T.E. (2000) Biochem. Cell Biol. 78:569.
Yu, T.W. and C.I. Bargmann (2001) Nature Neurosci. 4(Suppl):1169.
Stein, E. et al. (2001) Science 291:1976.
Koch, M. et al. (2000) J. Cell Biol. 151:221.
Yin, Y. et al. (2000) Mech. Dev. 96:115.
Livesey, F.J. (1999) Cell. Mol. Life Sci. 56:62.

Entrez Gene IDs

59277 (Human); 57764 (Mouse)

Alternate Names

beta-Netrin; hepar-derived netrin-like protein; netrin 4; Netrin4; Netrin-4; NTN4; PRO3091

Product Datasheets

Product Datasheet

COA

SDS

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 # 5,824,775, 6,218,526, and other U.S. and international patents pending.