Draxin

Manufactured by R&D Systems

Draxin is a recombinant protein that functions as an axon guidance molecule. It assists in the regulation of neuronal migration and patterning during development. The core function of Draxin is to facilitate the proper positioning of neurons within the nervous system.

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4 protocols using draxin

Draxin Binds to Netrin-1 In Vitro

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Example 11

Recombinant human Draxin was purchased from R&D systems. Biotinylated recombinant human UNC5B, Netrin1 and DCC were produced recombinantly using the described mammalian expression system (HEK293-6E). Biotinylated proteins were immobilised on the SA coated sensor chip and Draxin was injected sequentially in increasing concentrations (0 nM, 1.2 nM. 2.3 nM, 4.7 nM, 9.4 nM, 18.8 nM) for 3 min. Dissociation was allowed for 5 min in HBS-P. Binding was monitored and an interaction of Draxin to immobilised Netrin-1 was observed with a binding constant KD of approximately 20 to 100 nM. No binding of Draxin to UNC5B and DCC was detected (FIG. 14).

US10435442B2. Interaction of draxin and γ-netrins (2019-10-08). Max-Planck-Gesellschaft zur Foerderung der Wissenschaften e.V. [DE]. Inventors: Christian Söllner [DE], Xuefan Gao [DE], Christiane Nüsslein-Volhard [DE].

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Neurite Outgrowth Assay for MAP1B-/- Mice

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Newborn MAP1B-/- mice [22 (link)] of either sex and wild-type controls were sacrificed by decapitation. Cerebral cortices of newborn pups were dissected in ice cold Hank’s balanced salt solution (Gibco), 7 mM HEPES pH 7.3 (Gibco) and 2 mM L-glutamine (Gibco), embedded in matrigel (BD Biosciences) and cultured in neurobasal medium (Invitrogen) supplemented with B27 (Invitrogen), 2 mM L-glutamine and penicillin/streptomycin (50 units/ml; Invitrogen) in the presence or absence of draxin (10 nM; R&D Systems) or semaphorin 3A (100 ng/ml R&D Systems). After 48 h images of live explants were taken using an Axio-observer microscope (Zeiss) and the length of the longest neurite of each explant was measured as described [10 (link)] using image J software.

Meli R., Weisová P, & Propst F. (2015). Repulsive Axon Guidance by Draxin Is Mediated by Protein Kinase B (Akt), Glycogen Synthase Kinase-3β (GSK-3β) and Microtubule-Associated Protein 1B. PLoS ONE, 10(3), e0119524.

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Comprehensive Signaling Pathway Analysis

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HGF (R&D), Dkk1 (R&D), Wif1 (R&D), Draxin (R&D), sFRP1 (R&D), sFRP5 (R&D), BMP4 (R&D), Gremlin (R&D), Jagged 1 (Anaspec), BMP3 (R&D), BMP7 (R&D), Dll1 (R&D), Dll4 (R&D), DAPT (Sigma), DAPT-GSI (Sigma), TGF-α (R&D), Lrig1 (R&D), Shh (R&D), Ihh (R&D), Itraconazole (Sigma), Gant-61 (Tocris Bioscience), Calcitriol/Vit D (Sigma), Rapamycin (LC Laboratories), ICG-001 (Tocris), and Carnosic Acid (Sigma; also see Data S1)
Xenograft and in vivo Itraconazole dosing experiments: Beacon Pharmaceuticals (10 mg/ml).
Antibodies for IHC and FACS: AGR2 (Atlas, HPA007912), GDF15 (Atlas, HPA011191), β-catenin (BD Biosciences, 610154), and PTK7 (clone 188B; Miltenyi).

Buczacki S.J., Popova S., Biggs E., Koukorava C., Buzzelli J., Vermeulen L., Hazelwood L., Francies H., Garnett M.J, & Winton D.J. (2018). Itraconazole targets cell cycle heterogeneity in colorectal cancer. The Journal of Experimental Medicine, 215(7), 1891-1912.

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Draxin Interaction with Netrin-1

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Example 11

Recombinant human Draxin was purchased from R&D systems. Biotinylated recombinant human UNC5B, Netrin1 and DCC were produced recombinantly using the described mammalian expression system (HEK293-6E). Biotinylated proteins were immobilised on the SA coated sensor chip and Draxin was injected sequentially in increasing concentrations (0 nM, 1.2 nM. 2.3 nM, 4.7 nM, 9.4 nM, 18.8 nM) for 3 min. Dissociation was allowed for 5 min in HBS-P. Binding was monitored and an interaction of Draxin to immobilised Netrin-1 was observed with a binding constant KO of approximately 20 to 100 nM. No binding of Draxin to UNC5B and DCC was detected (FIG. 14).

US11505586B2. Interaction of Draxin and γ-netrins (2022-11-22). Max-Planck-Gesellschaft zur Foerderung der Wissenschaften e.V. [DE]. Inventors: Christian Söllner [DE], Xuefan Gao [DE], Christiane Nüsslein-Volhard [DE].

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