XE126 Rfz1myc CS2+
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||16798||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 4095
Vector typeXenopus Expression
Growth in Bacteria
Copy numberHigh Copy
SpeciesR. norvegicus (rat)
Insert Size (bp)2044
/ Fusion Protein
- Cloning method Restriction Enzyme
- 5′ sequencing primer SP6 (Common Sequencing Primers)
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.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:XE126 Rfz1myc CS2+ was a gift from Randall Moon (Addgene plasmid # 16798 ; http://n2t.net/addgene:16798 ; RRID:Addgene_16798)