Santa Cruz Biotechnology
Wnt3a Antibody
SC-136163
3A6
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We needed to determine whether recombinant Wnt3a would bind to heparan sulfate proteoglycans. We used a dotblot format where either PBS or proteoglycans were loaded onto nitrocellulose membranes. After a blocking step, purified Wnt3a was incubated with each sample well and later Santa Cruz Anti-Wnt3a was used to detect the growth factor signal.
Western Blot
Purified Recombinant Protein
1:100, overnight at 4 degrees Celsius
TBS, 0.05% Tween-20, 5% non-fat dry milk
Sheep anti-mouse HRP, 1:50000, 1hr, room temp
None
Enhanced Chemiluminescence
The expectation was that the Wnt3a growth factor would only bind to the proteoglycan samples and not the empty (blocked) wells. This turned out to be the case as the figure depicts. Increased amounts of proteoglycan lead to increased Wnt3a sginal (right side). On the left a pure growth factor standard curve was used.
N/A
Clean, low background, speciifc
High concentrations necessary
Great antibody, worked well in dot blots and western blots against purified, recombinant Wnt3a.