HaloTagged AP-2 σ2 was visualized by adding Janelia Fluor 646-HaloTag ligand, a gift from Luke Lavis30 . 100 nM ligand was added to cells and incubated at 37°C for 15 minutes. Cells were washed with fresh media and imaged immediately.
Visualization of HaloTagged AP-2 σ2 in SUM159 Cells
HaloTagged AP-2 σ2 was visualized by adding Janelia Fluor 646-HaloTag ligand, a gift from Luke Lavis30 . 100 nM ligand was added to cells and incubated at 37°C for 15 minutes. Cells were washed with fresh media and imaged immediately.
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Corresponding Organization :
Other organizations : The University of Texas at Austin, The University of Texas Health Science Center at San Antonio
Variable analysis
- Addition of HaloTag to both alleles of AP-2 σ2 in SUM159 cells
- Visualization of HaloTagged AP-2 σ2 using Janelia Fluor 646-HaloTag ligand
- Cell culture medium composition (1:1 DMEM high glucose:Ham's F-12, 5% fetal bovine serum, Pen/Strep/L-glutamine, 1 μg/ml hydrocortisone, 5 μg/ml insulin, 10 mM HEPES, pH 7.4)
- Cell seeding density (3x10^4 cells per coverslip)
- Transfection conditions (1-3 μg of plasmid DNA, 3 μL Fugene HD transfection reagent)
- Incubation conditions (37°C, 5% CO2)
- Staining protocol (100 nM Janelia Fluor 646-HaloTag ligand, 15 minutes incubation)
- No positive control mentioned.
- No negative control mentioned.
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