The Journal of Neuroscience, August 23, 2006, ():
Axon Guidance by Diffusible Chemoattractants: A Gradient of Netrin Protein in the Developing Spinal Cord
J. Neurosci. Kennedy et al.
26: 8866
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplemental Figure 1: Enhancement of netrin antigenicity.
Pan-netrin immunoreactivity detected in chick stage 23 brachial embryonic spinal cord. Adjacent sections showing netrin immunoreactivity with (A) and without (B) denaturing antigen enhancement (pan-netrin antibody PN-2). Cy3 coupled secondary antibody. Scale bar corresponds to 50 μm.
- supplemental material
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Supplemental Figure 2: Specificity of netrin immunoreactivity.
Stage 27 chick spinal cord illustrating the absence of immunostaining when antibody PN2 was preincubated for 1 hr with an ~3 fold molar excess of peptide 11760 (alkaline phosphatase coupled secondary antibody, BM Purple substrate, brightfield microscopy). Staining was similarly blocked by incubating antibody PN3 with an excess of recombinant netrin-1 VI-V protein, both on tissue sections and western blots. Application of the fluorescent secondary antibodies alone generated no staining in the embryonic spinal cord (data not shown).
