2S3P | 2S seed storage protein 3
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Arabidopsis thaliana dry seeds from wilde-type (WT) and mag2-2 mutant (defective in processing of the precursors) were homogenised in SDS sample buffer (100 mM Tris/HCl, pH 6.8, 4 % SDS w/v, 20 % glycerol v/v, 10 % 2-mercaptoethanol) and separated on SDS-PAGE and blotted to PVDF membrane in wet system. 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: 5000 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 recommendations.
Anti-2S3P antibodies are recognizing 2S albumin precursors (p2S) but not the mature forms (2S).
Alternative names: 2S albumin storage protein, NWMU2-2S albumin 3
Li et al. (2006). MAIGO2 is involved in exit of seed storage proteins from the endoplasmic reticulum in Arabidopsis thaliana. Plant Cell. 2006 Dec;18(12):3535-47. doi: 10.1105/tpc.106.046151.( Immunolocalisation by electron microscopy, Western blot, Arabidopsis thaliana)
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