Fd3 | Ferredoxin 3
AS20 4432 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Zea mays
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Product Information
Immunogen
Purified full length, tag cleaved, recombinant maize Fd3, UniProt: P27788
Host
Rabbit
Clonality
Polyclonal
Purity
Total IgG. Protein A purified in PBS, 50% glycerol. Filter sterilized.
Format
Liquid at 1 mg/ml.
Quantity
100 ĩg
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: 2000 - 1: 10 000 (WB)
Expected | apparent MW
16 kDa
Reactivity
Confirmed reactivity
Arabidopsis thaliana, Zea mays
Predicted reactivity
Brachypodium distachyon, Dichanthelium oligosanthes, Hordeum vulgare, Oryza sativa, Panicum hallii, Saccharum sp. , Setaria italica
Species of your interest not listed? Contact us
Species of your interest not listed? Contact us
Not reactive in
No confirmed exceptions from predicted reactivity are currently known
Application examples
Application examples
Fd3 purified from leaves of Zea mays, 50 ng was 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: 2000 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.
Total leaf extract of Arabidopsis thaliaa (1) and Zea mays (2), 10 µg 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: 10 000 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.
The antibody is also recognizing root-type ferredoxins expressed in leaves.
Different RNAi (lane 1-7) expressed in T1 Arabidopsis thaliana plants and wild type without any RNAi epxression. 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: 5000 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.
Fd3 purified from leaves of Zea mays, 50 ng was 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: 2000 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.
Total leaf extract of Arabidopsis thaliaa (1) and Zea mays (2), 10 µg 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: 10 000 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.
The antibody is also recognizing root-type ferredoxins expressed in leaves.
Different RNAi (lane 1-7) expressed in T1 Arabidopsis thaliana plants and wild type without any RNAi epxression. 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: 5000 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.
Additional information
Antibody reacts weakly with other ferredoxins: Arabidopsis thaliana and Zea mays Fd2 and Zea mays Fd6.
Background
Background
Ferredoxins are iron-sulfur proteins that transfer electrons in a wide variety of metabolic reactions. Fd3 is non-photosynthetic Fd expressed more in root than in leaf. Fd3 is localised in chloroplast and plastids.
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.
Hanke et al. (2003). A Post Genomic Characterization of Arabidopsis Ferredoxins. Plant Physiol. 134(1):255-64. doi: 10.1104/pp.103.032755.
Matsumura et al. (1997). A Nitrate-Inducible Ferredoxin in Maize Roots. Genomic Organization and Differential Expression of Two Nonphotosynthetic Ferredoxin Isoproteins. Plant Physiol. 114(2):653-60. doi: 10.1104/pp.114.2.653.
Hanke et al. (2003). A Post Genomic Characterization of Arabidopsis Ferredoxins. Plant Physiol. 134(1):255-64. doi: 10.1104/pp.103.032755.
Matsumura et al. (1997). A Nitrate-Inducible Ferredoxin in Maize Roots. Genomic Organization and Differential Expression of Two Nonphotosynthetic Ferredoxin Isoproteins. Plant Physiol. 114(2):653-60. doi: 10.1104/pp.114.2.653.
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