FER | Ferritin (plant)

AS10 674 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Cucumis stivus

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product information
Background		Ferritin is the major non-toxic iron storage protein complex in eukaryotic cells, consisting of 24 ferritin subunit polypeptides.
Immunogen		KLH-conjugated synthetic peptide derived from known plant ferritin sequences including Arabidopsis thaliana: ferritin-1 (chloroplastic), UniProt: Q39101, TAIR: AT5G01600; ferritin-2(expressed in roots), UniProt: Q9SRL5,TAIR: AT3G11050, ferritin-3 (chloroplastic), UniProt: Q9LYN2, TAIR: AT3G56090; ferritin-4 (chloroplastic), UniProt: Q9S756, TAIR: AT2G40300 (sequence identity 80 %)
Host		Rabbit
Clonality		Polyclonal
Clone
Purity		Serum
Format		Lyophilized
Quantity		200 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 tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications		Western blot (WB)
Related products		AS06 126 \| anti-Fer1 \| ferritin 1 (pre-apoferritin), algal antibody Plant protein extraction buffer Secondary antibodies
Additional information

application information
Recommended dilution		1 : 3000 (WB)
Expected \| apparent MW		28 \| 30 kDa
Confirmed reactivity		Arabidopsis thaliana, Cucumis stivus
Predicted reactivity		Hordeum vulgare, Musa sp., Oryza sativa, Phaseolus vulgaris, Physcomitrella patens, Solanum tuberosum, Ricinus communis,Triticum aestivum, Zea mays
Not reactive in		Lemna minor, Ostreococcus tauri
Additional information		Antibody is recognizing a chloroplastic form of plant ferritin
Selected references		Kovács et al. (2016). Revisiting the iron pools in cucumber roots: identification and localization. Planta. 2016 Jul;244(1):167-79. doi: 10.1007/s00425-016-2502-x. Epub 2016 Mar 22.

Application example

10 µg of total protein from Arabidopsis thaliala whole seedlings, various cellular fractions, extracted with (see below*) were separated on 15 % SDS-PAGE and blotted 1h to PVDF. Blots were blocked with 5% Non fat dairy milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 3000 overnight at 4°C with agitation. The antibody solution was decanted and the blot was washed four times for 5 min inPBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:15 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturers instructions. Exposure time was 180 seconds.

*Following is the protein extraction buffer composition that was used to extract the total protein from the whole seedlings:50mM Tris, 4% SDS, 1% PVPP , 5% Glycerol, 1mM PMSF, 2mM Pefabloc.

Courtesy of Anshika Jain and Erin Connolly, University of South Carolina, USA

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