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2020 (14-3-3/HCN1 C-terminus (HCN1 mutant expression/turnover (Cloning…
2020
14-3-3/HCN1 C-terminus
Reporter pulldown
100AA truncation reporters :check:
30AA truncation reporters :check:
site mutant reporters
10AA deletion :check:
S-A substitution
14-3-3 downregulation mechanism
Tissue pulldown
mutant 14-3-3 neg control
validate failed binding with p-53 antibody
co-IP
Brain
HEK293T cells
HCN1 mutant expression/turnover
Cloning AAVS1-CAG: SNAP-HCN1
Single mutants :check:
Double mutants
triple mutants :check:
WT :check:
S->A mutant (Genscript) :green_cross:
Stable lines genration
WB expression of all mutants
SNAP-tag pulse-chase
HCN domain (weekend project)
Cloning
Clover
WT :check:
VSL :fire:
HCNd :fire:
untagged
WT :check:
HCNd :check:
VSL :check:
mCherry
WT :check:
assembly
co-IP
BN-PAGE
trafficking
trafficking rescue
external ER export signal?
HCN1/vision
ERG
Dark adapted single flash
rod
cone N=3
flicker
rod (N=1)
cone (N=6)
photopic fflash
rod (N=1)
cone (N=6)
Light aversion
Naive
cone (N=10)
rod (N=1)
adapted
OMR (solessio)
IHC (cone arrestin + Cre + HCN1)
Cone
Rod
Kv8.2/Kv2.1 stoichiometry
cloning FRET pairs
mClover-Kv2.1 :check:
mRuby-Kv2.1 :check:
mClover-Kv8.2
mRuby-Kv8.2 :check:
FRET experiment
T1 domain (X-ray core)
Kv2.1 T1-TEV cloning :check:
Kv8.2 T1-TEV cloning :check:
Cav Western blotting
a1f KO WB set
a2d4 WB set