Native-PAGE Analysis of BiP Oligomers
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Corresponding Organization : University of Cambridge
Variable analysis
- Type of gel used (4.5% stacking gel and a 7.5% separation gel)
- Voltage used for separation (120 V)
- Duration of separation (2 hours)
- Type of buffer used for separation (Tris-glycine running buffer)
- Type of buffer used for transfer (Tris-glycine blotting buffer with 0.04% (w/v) SDS)
- Duration of transfer (16 hours at 30 V)
- Concentration of purified BiP protein loaded (7 µg per lane)
- Amount of total protein from cell lysates loaded (30 µg per lane)
- Separation of purified BiP proteins or proteins from mammalian cell lysates to detect BiP oligomers
- Visualization of separated proteins by Coomassie staining or immunodetection on PVDF membrane
- PH of running buffer (~8.8)
- PH of blotting buffer (~9.2)
- Methanol concentration in blotting buffer (20% v/v)
- Cell line used (CHO-K1)
- Positive control: Purified BiP protein
- Negative control: Not explicitly mentioned
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