IKP

Anti-Fd1 | Ferredoxin 1 (chloroplastic)

Product no: AS20 4434
AS20 4434 | Clonality: Polyclonal  |  Host: Rabbit |  Reactivity: Arabidopsis thaliana, Zea mays
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  • Product Info
  • Immunogen: Purified full length, tag cleaved, recombinant maize Fd1, UniProt: P27787
    Host: Rabbit
    Clonality: Polyclonal
    Purity: Total IgG. Protein A purified in PBS, 50% glycerol. Filter sterilized.
    Format: Liquid at 1 mg/ml.
    Quantity: 100 µg
    Storage: Store 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: ELISA (ELISA), Western blot (WB)
    Recommended dilution: 1: 1000 - 1: 5000 (WB)
    Expected | apparent MW: 12 | 15 kDa
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana, Zea mays
    Predicted reactivity: Brassica napus, Brassica oleracea, Eutrema salsugineum, Raphanus sativus
    Species of your interest not listed? Contact us
    Not reactive in: Nicotiana benthamiana
  • Application Examples
  • Western blot using anti-Fd1 antibodies
    Recombinant Fd1 from Zea mays (1), 10 μg/well of leaf total protein of Arabidopsis thaliana wild type leaf (2), Zea mays leaf (3) were freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE. For IP, 150mM NaCL, 1% Triton X-100, 50 mM Tris-HCl (pH 8.0) and denatured with 4X SDS buffer at 95°C for 5 min. Samples were separated on 10% SDS-PAGE and blotted 1h to PVDF membrane. 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: 1000 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 recommendation.

    Molecular weight of mature Fd1 is around 12 kDa but it migrates at 15 kDa in applied gel system. 


    Western blot using anti-Fd1 (plant) antibodies

    200 nmol of recombinant Ferredoxin-1 (Fd1) from Arabidopsis thaliana (1),
    200 nmol of recombinant Ferredoxin-2 (Fd2) from Arabidopsis thaliana (2),
    200 nmol of recombinant Ferredoxin-3 (Fd3) from Arabidopsis thaliana (3),
    20 nmol of recombinant Ferredoxin-4 (Fd4) from Arabidopsis thaliana (4),
    Leaf extract of Arabidopsis thaliana, soluble fraction precipitated with 70 % ammonium sulfate (5),
    Leaf extract of Arabidopsis thaliana, insoluble fraction precipitated with 70 % ammonium sulfate (6), 
    Root extract of Arabidopsis thaliana (7).  Samples were separated on 10% SDS-PAGE and blotted 1h to PVDF membrane. 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: 200 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 recommendation.
  • Background
  • Background: Ferredoxins are iron-sulfur proteins that transfer electrons in a wide variety of metabolic reactions. It occupies a key position both for transferring the photoreducing power to Fd-NADP+ oxidoreductase (FNR), hence the formation of NADPH, and for mediating the cyclic electron flow around photosystem I (PSI).
  • Product Citations
  • Selected references: Hanke and Hase (2008). Variable Photosynthetic Roles of Two Leaf-Type Ferredoxins in Arabidopsis, as Revealed by RNA Interference. Photochem Photobiol. 84(6):1302-9. doi: 10.1111/j.1751-1097.2008.00411.x.
    Kimata and Hase (1989). Localization of Ferredoxin Isoproteins in Mesophyll and Bundle Sheath Cells in Maize Leaf. Plant Physiol. 89(4):1193-7. doi: 10.1104/pp.89.4.1193.
  • 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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