To explore the effect of specific inhibitors on axonal growth, 6-well plates were coated with 0.01% PLL overnight. The next day, they were washed three times with PBS and dried at 37°C. Then, 3-µl droplets of CSPGs (50 µg/ml; EMD Millipore) were spotted onto the 6-well plates for 4 h at 37°C. After the droplets dried, the six-well plates were covered with laminin(LN) (10 µg/ml; Invitrogen) for 2 h at 37°C. They were then washed three times with PBS and stored at 37°C before neuron plating.
Laminin ln
Manufactured by Thermo Fisher Scientific
Laminin (LN) is a glycoprotein found in the extracellular matrix of many tissues. It plays a key role in cell adhesion, migration, and differentiation. LN serves as a structural component and provides a surface for cell attachment.
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Lab products found in correlation
2 protocols using laminin ln
1
Investigating Axonal Growth Inhibitors
2
Micropatterned Substrate Fabrication
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Micropatterned 2D substrates were produced as described elsewhere59 (link). In brief, the pattern was transferred from a silicone (Polydimethylsiloxane (PDMS), Sylgard 184, Dow Corning, #(400)000105989377) stamp onto a titanium-gold (Ti: 2 nm, Au: 20 nm thickness) sputtered cover slip. The pattern consisted of octadecylmercaptan (ODM, 1.5 mM in ethanol, Aldrich Chemistry, #01858-25ML). Remaining regions were passivated with HS-C11-(EG)6-OH (0.1 mM in ethanol, Prochimia, #TH-001-m11.n6-0.01). For protein coating, fibronectin (FN, Sigma-Aldrich #F2006) or laminin (LN, Invitrogen, #23017-015) was diluted in PBS and used in a final concentration of 10 µg/ml. Incubation occurred for 30 min at RT. As a control, scaffolds were coated with Poly-d -Lysine (PDL, Gibco, #A38904-01) in a concentration of 50 µg/ml diluted in ddH2O for one hour at RT. The proteins adsorb to the ODM-covered regions but not to the HS-C11-(EG)6-OH-covered regions, which allows patterning of the substrate.
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