VDE | Violaxanthin de-epoxidase

Product no: AS15 3091

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

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  • Product Info
  • 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 l
    Reconstitution: For reconstitution add 200 l of sterile water
    Storage: Store lyophilized/reconstituted at -20C; 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
  • 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

    Application examples:

    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
  • Additional information (application):

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

  • 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.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Posters Collection
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Buy 2 items of this product for 226.00 €/items
Buy 3 items of this product for 205.00 €/items
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