AS20 4405 Anti-2S3P | 2S seed storage protein 3

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).

Anti-2S3P antibodies are recognizing 2S albumin precursors (p2S) but not the mature forms (2S)

2S seed storage protein 3, one of major seed storage proteins is synthesized on the endoplasmic reticulum as precursor and then transported to storage vacuoles, where it is processed by an asparaginyl endopeptidase to produce two mature polypeptides referred to as large and small subunits which are linked by disulfide bonds. Subcellular localisation: vacuole.

Alternative names: 2S albumin storage protein, NWMU2-2S albumin 3

1. Shirakawa et al. (2014). CONTINUOUS VASCULAR RING (COV1) is a trans-Golgi network-localized membrane protein required for Golgi morphology and vacuolar protein sorting. Plant Cell Physiol. 2014 Apr;55(4):764-72. doi: 10.1093/pcp/pct195. (Western blot, Arabidopsis thaliana)
2. 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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