MEB2 | Membrane protein of ER body 2
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Arabidopsis thaliana 7 day seedling crude extract was freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE and denatured with 4X SDS buffer at 95°C for 5 min. Protein load/well is 10 µg. Sample was separated on 7.5 % SDS-PAGE and blotted at 15V overnight using wet transfer to PVDF membrane. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10 000 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendation.
Calculated MW of MEB2 is 61 kDa, while apparent MW appears to be 82 kDa (ER membrane protein).
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