Arf1 | ADP-ribosylation factor 1
AS08 325 | clonality: polyclonal | host: rabbit | reactivity: A.thaliana, Elaeis p.,L. longiflorum, N.tabacum, O. sativa, P. patens, Ch.reinardtii | cellular [compartment marker] of Golgi in immunolocalization and COP1 in western blot
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1:1000 with standard ECL (WB), 1: 1000 (IF). 1: 100 (IG)
|Expected | apparent MW||
21 kDa (Arabidopsis thaliana)
Arabidopsis thaliana, Chlamydomonas reinhardtii, Elaeis sp. , Lilium longiflorum, Nicotiana tabacum, Oryza sativa, Petunia hybrida, Physcomitrella patens
B. juncea, B.napus, C.annum, Cucumis sp., D.carota, E.guineensis,G.max, H.orientalis, M.truncatula, N.benthamina, S.tuberosum, C.rubella, H. vulgare, P. trichocarpa, Zea Mays,
|Not reactive in||
References describing immunolocalization (IF) and (IG) studies:Pimpl et al (2000). In Situ Localization and in Vitro Induction of Plant COPI-Coated Vesicles. Plant Cell. 2000 Nov;12(11):2219-36.
Ritzenthaler et al. (2002). Reevaluation of the Effects of Brefeldin A on Plant Cells Using Tobacco Bright Yellow 2 Cells Expressing Golgi-Targeted Green Fluorescent Protein and COPI Antisera. Plant Cell. 2002 Jan;14(1):237-61.
|Selected references||Marais et al. (2015). The Qb-SNARE Memb11 interacts specifically with Arf1 in the Golgi apparatus of Arabidopsis thaliana. J Exp Bot. 2015 Jul 24. pii: erv373.
Wang et al. (2015). UDP-D-galactose synthesis by UDP-glucose 4-epimerase 4 is required for organization of the trans-Golgi network/early endosome in Arabidopsis thaliana root epidermal cells. J. Plant Res. 2015 May 27. (immunogold application)
Komsic-Buchmann et al. (2014). The Contractile Vacuole as a Key Regulator of Cellular Water Flow in Chlamydomonas reinhardtii. Eukaryotic cell (13), Issue 11: 1421-30.
Frescatada-Rosa et al. (2014). High Lipid Order of Arabidopsis Cell Plate Membranes Mediated by Sterol and DYNAMIN-RELATED PROTEIN1A Function. Plant J. 2014 Sep 18. doi: 10.1111/tpj.12674.
Ranocha et al. (2013). Arabidopsis WAT1 is a vacuolar auxin transport facilitator required for auxin homoeostasis. Nat Commun. 2013;4:2625. doi: 10.1038/ncomms3625.
Liu et al. (2013). Brittle Culm1, a COBRA-Like Protein, Functions in Cellulose Assembly through Binding Cellulose Microfibrils. PLoS Genet 9(8): e1003704. doi:10.1371/journal.pgen.1003704 (Oryza sativa, western blot)
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 (recommended secondary antibody AS09 602). 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%.
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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