IKP

Anti-Fd2 | Ferredoxin 2 (chloroplastic)

Product no: AS20 4433
AS20 4433 | Clonality: Polyclonal  |  Host: Rabbit |  Reactivity: Arabidopsis thaliana, Zea mays
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  • Product Info
  • Immunogen: Purified full length, tag cleaved, recombinant maize Fd2, UniProt: P16972
    Host: Rabbit
    Clonality: Polyclonal
    Purity: Total IgG. Protein A purified in PBS, 50% glycerol. Filter sterilized.
    Format: Liquid at 2 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: 15 kDa
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana, Synechocystis PCC 6803, Zea mays
    Predicted reactivity: Brachypodium distachyon, Oryza sativa, Setaria italica, Sorghum bicolor
    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Western blot using anti-Fd2 (plant) antibodies

    10 μg/well of Arabidopsis thaliana total leaf extract (1),  10 μg/well of Zea mays total leafextract (2) 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.

    Western blot with anti-Fd2 antibodies on cyanobacterial sample

    5 μg ofcrude extract of Synechocystis PCC6803 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.
  • Background
  • Background: Ferredoxins are iron-sulfur proteins that transfer electrons in a wide variety of metabolic reactions. 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). Fd2 is most abundant Fd isoprotein expressed in plant leaves.
  • Product Citations
  • Selected references: Ramirez et al. (2013). Glutathione and ascorbic acid protect Arabidopsis plants against detrimental effects of iron deficiency.
    Hanke et al. (2004). A post genomic characterizationof Arabidopsis ferredoxins. Plant Physiol. 2004 Jan;134(1):255-64. Epub 2003 Dec 18 (Western blot, Arabidopsis thaliana)
  • 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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