Electrophoretic mobility shift assays (EMSAs) were used to assess the regulation of Oct-4/Sox2 by CDX2. Probes with the CDX2-binding site sequence (see Table 1 ) were labeled with Digoxigenin-11-deoxyuridine 5-triphosphate (DIG-ddUTP). Cell protein-DNA complexes were examined with and without competitors or the anti-CDX2 antibody. After electrophoretic separation, the complexes were probed auto-radiographically using a 2nd-generation DIG Gel Shift Kit (Roche, Mannheim, Germany).
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Investigating CDX2 Regulation of Oct-4/Sox2 using EMSA
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Anti antibody
Binding site
Cell
Ddutp
Digoxigenin 11 deoxyuridine triphosphate
Electrophoretic
Electrophoretic mobility shift assays
Oct 4
Protein dna
Sox2
Corresponding Organization :
Other organizations : Stomatology Hospital, Sun Yat-sen University
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Variable analysis
independent variables
- Presence or absence of competitors
- Presence or absence of anti-CDX2 antibody
- Electrophoretic mobility of protein-DNA complexes
- CDX2-binding site sequence of the probe
- Labeling of the probe with Digoxigenin-11-deoxyuridine 5-triphosphate (DIG-ddUTP)
- Electrophoretic separation of the protein-DNA complexes
- Probing of the complexes using a 2nd-generation DIG Gel Shift Kit (Roche, Mannheim, Germany)
- Positive control: Protein-DNA complexes without any competitors or antibody
- Negative control: Not explicitly mentioned
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