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URL: http://www.addgene.org/16798
Proper Citation: RRID:Addgene_16798
Insert Name: Rfz1
Organism: Rattus norvegicus
Bacterial Resistance: Ampicillin
Vector Backbone Description: Backbone Size:4095; Vector Backbone:pCS2+; Vector Types:Xenopus Expression; Bacterial Resistance:Ampicillin
Comments: Constructed by: Julia Yang-Snyder (subclone), Jeff Brown (tags), 4/19/96. Rfz1 from Henk Roelink. This sequence has a few differences from the published sequence. Sites listed in Rfz1 are not a complete list. MYC tag version made by Jeff Brown (2/98-3/98), inserted the epitope into the StyI site (1904). The MYC epitope has a BglII site in it. The 3' Sty site is destroyed by the tags, but the 5' site is intact. Epitope can be detected in PBS/Formaldehyde fixed tissue permeabilized with 0.2% Triton. The Myc tagged Rfz1 has been IP'd from a TBS/1% Triton buffer. Frizzled tends to ppt out of SDS sample buffer when boiled, so either warm samples to ~70 degrees Celsius for a few minutes before loading samples or use a strong denaturing loading buffer and incubate the samples at room temp for half an hour before loading. The StyI site is in the 3rd extracellular loop of the protein.
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Source: Addgene
