The ARF1 protein is localized to the Golgi apparatus and has a central role in intra-Golgi transport. It is a small GTPase that undergoes a GDP/GTP nucleotide exchange cycle and it is an important regulator of cellular trafficking.
immunogen
recombinant GST fusion of full length of Arabidopsis thaliana ARF1 At2g47170
antibody format
rabbit
polyclonal
serum
lyophilized
quantity
200 µl
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.
dicots including B. juncea, B.napus, C.annum, D.carota, E.guineensis,G.max, H.orientalis, M.truncatula, N.benthamina, S.tuberosum monocots including O.sativa, moss Physcomitrella patens
not reactive in
no confirmed exceptions from predicted reactivity known in the moment
additional information
to be added when available
selected references
Pertl et al. (2009). The pollen organelle membrane proteome reveals highly spatial-temporal dynamics during germination and tube growth of lily pollen. j. Proteome Res. 11:5142.5152.
application example
50 µg of total protein from (1) Nicotiana tabacum protoplast total protein, (2) Arabidopsis thaliana protoplast soluble protein, (3) Arabidopsis thaliana protoplast total protein were separated on 10 %SDS-PAGE and blotted 2h to nitrocellulose (Semi-dry, 200mA). Filters were blocked over night with 5% low-fat milk powder in TBS and probed with anti-Sec21p antibodies (AS08 327, 1:1000, 1h) and secondary anti-rabbit (1:20000, 1 h) antibody (HRP) in TBS-Tween. Signal was detected with standard ECL andexposure time for this image was 1 minute.
Protoplasts were extracted in 50mM Tris, 10 mM EDTA and Triton X100, 0.02%.
Immunofluorescence
Specificity testing of rabbit anti-ARF1 serum. Immunofluorescence labelling of rabbit anti-ARF1 antibody (red) in 5-day-old root epidermal cells of the Arabidopsis thaliana ecotype Columbia-0 (WT) or seedlings expressing the ADP-RIBOSYLATION FACTOR 1 (AtARFA1c; accession At2g47170) fused to EGFP (green) (Xu, J. and Scheres, B. 2005. Plant Cell 17, 525-536). The rabbit anti-ARF1 antibody was diluted 1:1000 and the secondary antibody, donkey anti-rabbit CY5-coupled (Jackson ImmunoResearch) was diluted 1:300. The nuclei were stained with DAPI (blue). Note the co-labelling of ARF1-GFP with the anti-ARF1 antibody (arrowheads) and the additional labelling (potentially of other ARF1 variants) by the anti-ARF1 antibody (arrows). The antibody staining permeability was limited to the 1-2 outermost layers of the whole-mounted root tips.
Courtesy of Dr. Anna Gustavsson and Dr. Markus Grebe
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