Embryos were injected and electroporated with an EGFP plasmid (pMES) at HH St 10 and incubated for 72 hours. Embryos were then sectioned transversely, using a feather blade (~150um thick), between the fore- and hindlimbs and placed on a Millicell culture insert (EMD Millipore, Billerica, MA). BDNF- (Peprotech, Rocky Hill, NJ) or PBS- soaked Affigel beads (Bio-Rad, Hercules, CA) were placed in the slices along the dorsal migratory path of neural crest cells but in ectopic locations that the cells normally avoid (i.e. adjacent to notochord, ventral to spinal nerve, dorsal aorta), using fine glass pipettes and tungsten needles. The Millipore filter was then placed in a glass bottom petri dish and tissue time-lapse imaged. For function blocking experiments, trunk transverse tissue sections were cultured as above and the following function blocking antibodies were added to the culture media: anti-BDNF (1:500; Millipore, #AB1513P), anti-trkB (1:500; R and D systems, #AF1494) or CHEX, anti-p75 (1:1,000; kind gift of F. Lefcort/L. Reichardt, MSU).
Ab1513p
Manufactured by R&D Systems
The AB1513P is a laboratory equipment product. It is designed for general laboratory use. The core function of the AB1513P is to perform specific laboratory tasks. However, a detailed and unbiased description of its features and capabilities is not available at this time.
Automatically generated - may contain errors
2 protocols using ab1513p
1
Investigating Neural Crest Cell Migration
2
Investigating Neural Crest Cell Migration Using BDNF Stimulation
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Embryos were injected and electroporated with an EGFP plasmid (pMES) at HH St. 10 and incubated for 72 h. Embryos were then sectioned transversely, using a feather blade (∼150-μm thick), between the fore- and hindlimbs and placed on a Millicell culture insert (EMD Millipore, Billerica, MA). BDNF- (Peprotech, Rocky Hill, NJ) or PBS-soaked Affigel beads (Bio-Rad, Hercules, CA) were placed in the slices along the dorsal migratory path of neural crest cells but in ectopic locations that the cells normally avoid (that is, adjacent to notochord, ventral to spinal nerve, dorsal aorta), using fine glass pipettes and tungsten needles. The Millipore filter was then placed in a glass bottom Petri dish and tissue time-lapse imaged. For function-blocking experiments, trunk transverse tissue sections were cultured as above and the following function blocking antibodies were added to the culture media: anti-BDNF (1:500; Millipore, #AB1513P) or anti-trkB (1:500; R and D systems, #AF1494).
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