Immunostaining Procedure for Microtubule Analysis
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Corresponding Organization :
Other organizations : Institut Jacques Monod, Université Paris Cité, Centre National de la Recherche Scientifique
Variable analysis
- Ciliobrevin D
- Microtubules (MT) staining
- Fixation method (in bulk, and under microscope)
- Fixation duration (70 min)
- Fixation solution (100 mM Hepes, pH 6.9, 50 mM EGTA, 10 mM MgSO4, 2% formaldehyde, 0.2% glutaraldehyde, 0.2% Triton X-100, and 400 mM glucose)
- Rinsing procedure (3 times for 10 min in PBS plus Tween 20, and 1 time in PBS)
- NaBH4 treatment (0.1% in PBS for 30 min)
- Blocking procedure (PBT plus 5% goat serum and 0.1% BSA for 30 min)
- Primary antibody (anti–α-tubulin, clone DM 1A, 1/8,000, 48 h)
- Secondary antibody (fluorophore-conjugated anti-mouse, 1/750, 4 h)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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