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URL: http://www.addgene.org/16797
Proper Citation: RRID:Addgene_16797
Insert Name: Rfz1-HA-CS2+
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. Comments: Rfz1 from Henk Roelink. This sequence has a few differences. from the published sequence. Sites listed in Rfz1 are not a complete list. and I haven't searched through the CS2+ sequence to see if/where those sites. are present. HA tag version made by Jeff Brown (2/98-3/98) insert the epitope. into the StyI site (1904). The 3' Sty site is destroyed by the tags, but. the 5' site is inteact. Epitope can be detected in PBS/Formaldehyde fixed. tissue permeablized with 0.2% Triton. 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 and hour before loading. The StyI site is in the 3rd extracellular loop of the protein.
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Source: Addgene
