Anti-dVPE (At) antibody, rabbit polyclonal
Product code81-118Size

100 µgStorage-20℃

Concentration

2.0 mg/ml

Buffer

PBS- with 50% glycerol

PurityPurified IgG fraction with protein A from rabbit antiserum.

Immunogen

Recombinant His6-dVPE (amino acids 40-407) of Arabidosis thaliana.

IsotypeRabbit IgG

Reactivity

Arabidopsis thaliana. Not tested in other species.

Special notes

Subcellular localization: Seed specific. Restricted to developing seeds at 7 days
after anthesis, and, at lower levels, detected in flowers siliques, specifically inseed coats.

Application

1. Western blotting (1/5,000)
2. Immunofluorescent staining (1/500)
3. Immunoelectron microscopy (1/5,000)

Backgroundδ VPE (Vacuplar-Processing Enzyme delta-isozyme) is Asparagine specificendopeptidase that may be involved in processing of proteins targeted tovacuoles (By similarity). Probably involved in post-translational proteolysis ofseed storage proteins in the protein storage vacuole of developing seeds(PubMed:12417707, PubMed:14688293). Exhibits a caspase-1-like activity inextracellular granules. At the early stage of seed development, required for theformation of the seed coat, by regulating cell death of specific cell layers in innerintegument (PubMed:15705955).Length:466 amino acide. Mass; 52,085. Signal
peptide of 24 amino acids is removed in mature protein.

Data Link

Data Link: UniProtKB Q9LJX8 (VPED_ARATH)

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.

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Data Images: 81-118 Anti-dVPE (At) antibody, rabbit polyclonal

Fig.1 Western blot of dVPE in extract of maturing siliques of arabidopsis
Crude extract of maturing siliques of Arabidopsis thaliana was run on SDS-PAGE (15-20% gradientgel) and blotted to PVDF membrane by wet system. Blocking was done with 3% skim milk. The antiPYK10 (intermial) antibody was used at 0.4 μg/ml. Secondary antibody (goat anti-rabbit IgG antibody
HRP-conjugated, ab97051) was used at 1/10,000 dilution. dVPE is produced as an inactive precursor
with 52 kDa mass and processed into mature active form with 37 -38 kDa mass (Ref 1)

Fig 2 Immunofluorescence analysis of surfaces of the inner integuments with anti-dVPE antibodies
Left: Differential Interference Contrast image.
Right: Immunofluorescent staining.
The developing seeds of Arabidopsis that had walking-stick-shaped embryos were fixed for 40 min in
7.2% (w/v) formaldehyde, 0.1% (v/v) Nonidet P-40, 10% (v/v) dimethyl sulfoxide, and 50 mM Naphosphate buffer, pH 7.2, washed twice with TBS-T for 5 min, incubated in TBS-T containing 5% (w/v)
Cellulase Onozuka R-10 and 2% (w/v) Pectolyase Y-23 for 20 min at 30C, washed twice with TBS-T,incubated in blocking buffer (2% BSA and TBS-T) for 30 min, incubated with anti-dVPE antibodies(diluted 500-fold) in the blocking buffer for 40 min, washed three times for 5 min each, incubated for
1 h with goat anti-rabbit IgG antibodies conjugated with Alexa Fluor 488, washed three times for 5min with TBS-T, and mounted
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Reference. This antibody was described in Ref.1 and used in the following publications.
1. Nakaune S et al. A vacuolar processing enzyme, deltaVPE, is involved in seed coat formation at theearly stage of seed development. Plant Cell. 2005 Mar;17(3):876-87.
PMID: 15705955.WB, IF, Immunoelectron microscopy (arabidopsis)
2. Kunieda T et al. NAC family proteins NARS1/NAC2 and NARS2/NAM in the outer integument
regulate embryogenesis in Arabidopsis. Plant Cell. 2008 Oct;20(10):2631-42. PMID: 18849494 WB(arabidopsis)
3. Kunieda T et al. Spatiotemporal secretion of PEROXIDASE36 is required for seed coat mucilage
extrusion in Arabidopsis. Plant Cell. 2013 Apr;25(4):1355-67. PMID: 23572548.WB (arabidopsis)

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