block + ab

title: block + ab updated: 2024-02-02T18:16:31 created: 2024-01-30T17:26:59

Blocking

• PBTA = PBS, 0.25% TRITON, 3 % BSA, 0.01% NaN3
• 45 min RT
• 200ul/well Antibodies
• FLAG tag 1:100 in 3% BSA in PBST
• HA tag 1:500 --...
• 200 ul/well
• Overnight 4 °C

Wash PBST 3x Secondary antibodies

• Donkey a-mouse 488 (dil 1:2000)
• Donkey a-rabbit 488 (dil 1:2000)
• Donkey a-rabbit 594 (dil 1:1000)
• 200 ul/well, 1 h RT

Hoechst 0,1 ug/ml Mounting

Samples:

| Slide | 1 | 2 | 3 | | --- | --- | --- | --- | | . | Notch mito ✓ | VDAC mito ✓ | GSK mito ✓ | | .. | Notch + Vdac ✓ | Notch + gsk ✓ | Vdac + gsk ✓ | | ... | N + ( G + V ) mito✓ | (Notch) + Vdac mito ✓ | (Notch) + GSK mito ✓ |

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