XE127 Rfz1HA-CS2+
(Plasmid #16797)

• Depositing Lab: Randall Moon
• Publication: Xenopus Expression Plasmids (unpublished)

Backbone

• Vector backbone: pCS2+
• Backbone size w/o insert (bp): 4095
• Vector type: Xenopus Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Rfz1-HA-CS2+
• Alt name: fzd1
• Alt name: frizzled
• Species: R. norvegicus (rat)
• Insert Size (bp): 2044
• Entrez Gene: Fzd1
• Tag / Fusion Protein:
  • HA (internal)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: SP6

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor 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.