VDE | Violaxanthin de-epoxidase

AS15 3091 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Brachypodium distachyon, Deschampsia antarctica, Oryza sativa

Product Information

Immunogen

His-tagged, recombinant VDE from Arabidopsis thaliana, overexpressed in E.coli, Uniprot:Q39249 , TAIR: AT1G08550

Host Rabbit

Clonality Polyclonal

Purity Serum

Format Lyophilized

Quantity 50 µl

Reconstitution For reconstitution add 200 µl of sterile water

Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.

Tested applications Western blot (WB)

Recommended dilution 1 : 2000 (WB)

Expected | apparent MW

Propeptide 52 kDa, mature protein: 39.7 kDa, migrates at 40 kDa (Arabidopsis thaliana)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Brachypodium distachyon, Deschampsia antarctica, Oryza sativa,

Predicted reactivity Brassica napus, Camellia sinensis, Citrus limon, Cucumis sativus, Chrysanthemum morifolium, Citrus sinensis, Coffea arabica, Fragaria ananassa, Glycine soja, Gossypium arboreum, Lactuca sativa, Lycium barbarum, Morus notabilis, Nicotiana tabacum, Phyllostachys edulis, Picea abies, Prunus humilis, Spinacia oleracea, Solanum lycopersicum, Theobroma cacao, Triticum aestivum, Zingiber officinale, Vitis vinifera
Species of your interest not listed? Contact us

Not reactive in diatoms

Application examples

Application examples Western blot using anti-VDE antibodies in Arabidopsis, Brachypodium and rice

Samples:
R – 10 µL of total protein extract of Oryza sativa (50 mg macerated leaves in 100 µL Lysis Buffer).
B – 10 µL of total protein extract of Brachypodium distachyon (50 mg macerated leaves in 100 µL Lysis Buffer).
A – 10 µL of total protein extract of Arabidopsis thaliana diluted to 1:2 (50 mg macerated leaves in 200 µL 2X Laemmli Buffer). MW marker: NZYColour Protein Marker II from NZYTech.

10 µL of total protein extracted freshly from Arabidopsis thaliana, Brachypodium distachyon, and Oryza sativa with Lysis Buffer (7M Urea, 2M Thiourea, 4% CHAPS, 35 mM Tris), and denatured in 1X Laemmli with 25 mM DTT at 95ºC for 4 min, were separated on 10 % SDS-PAGE and blotted 1h to PVDF membrane (pore size of 0.45 μm, GE Healthcare), using wet transfer. Blot was blocked with Blocking solution containing 5% milk in TBS-T, for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:2000 in Blocking solution containing 5% milk in TBS-T, for O/N at 4ºC, with agitation. The antibody solution was decanted, and the blot was washed 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG, horse radish peroxidase conjugated) diluted to 1:20 000 in Blocking solution containing 5% milk in TBS-T, for 1h/RT with agitation. The blot was washed 6 times for 5 min in TBS-T at RT with agitation and developed for 30 seconds with Agrisera ECLSuperBright. Exposure time was 10 seconds.

Courtesy of Dr. Nelson Saibo,Plant Gene Regulation Lab (GPlantS Unit), ITQB NOVA, Portugal

Reactant: Solanum lycopersicum (Tomato)

Application: Western Blotting

Pudmed ID: 34439953

Journal: Biology (Basel)

Figure Number: 9A

Published Date: 2021-07-28

First Author: Trojak, M. & Skowron, E.

Impact Factor: None

Open Publication

Western Blot analyses (a) and densitometric analyses of PsbS (b), VDE (c), PGRL1 (d), and cytf (e) proteins in leaves of tomato plants (Solanum lycopersicum L. cv. Malinowy Ozarowski) grown under different light conditions (see Material and Methods for details). The bands were normalized to the appropriate ? subunit of ATP synthase (ATPB) band (loading control, Figure S4) (a). The bar diagrams (b–e) represent pixel volumes (densitometric analyses) of proteins in samples. Each value represents the mean ± SD (n = 3) considering the control sample (WL) value as 1 (100%). Different letters indicate significant differences between treatments (p = 0.05) with a Tukey’s HSD test. MW—molecular weight.

Additional information

Extraction buffer must contain 7 M urea for Brachypodium distachyon and Oryza sativa samples.
This product can be sold containing ProClin if requested.

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Background

Background VDE (violaxanthin de-epoxidase) is a part of the xanthophyll (or violaxanthin) cycle for controlling the concentration of zeaxanthin in chloroplasts. This enzyme is catalyzing the two-step mono de-epoxidation reaction. Zeaxanthin induces the dissipation of excitation energy in the chlorophyll of the light-harvesting protein complex of photosystem II.