ARF1 | ADP-ribosylation factor 1

AS08 325 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, C.reinhardtii, Elaeis sp. , L. longiflorum, M. truncatula, N. tabacum, O. sativa, P. hybrida cv Mitchell, P. patens, S. tuberosum | Cellular [compartment marker] of Golgi in immunolocalization and COP1 in Western blot

Product Information

Immunogen

Recombinant GST fusion of full length of Arabidopsis thaliana ARF1 (P36397, AT2G47170)

Host Rabbit

Clonality Polyclonal

Purity Serum

Format Lyophilized

Quantity 50 µl

Reconstitution For reconstitution add 50 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 Immunofluorescence (IF), Immunogold (IG), Western blot (WB)

Recommended dilution 1 : 1000 (IF). 1 : 100 (IG), 1 : 1000 (WB)

Expected | apparent MW

21 kDa (Arabidopsis thaliana)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Chlamydomonas reinhardtii, Elaeis sp. , Lilium longiflorum, Medicago truncatula, Nicotiana tabacum, Oryza sativa, Petunia hybrida cv Mitchell, Physcomitrella patens, Solanum tuberosum

Predicted reactivity

Brassica juncea, Brassica napus, Capsella rubella, Capsicum annum, Cucumis sp., Daucus carota, Elaeis guineensis, Glycine max, Helleborus orientalis, Hordeum vulgare, Medicago truncatula, Nicotiana benthamina, Ostreococcus tauri , Populus trichocarpa, Zea mays

Not reactive in

Microsporidia sp.

Application examples

Application example

Western blot detection using Arf1 antibody

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 chemiluminescence detection reagent and exposure time for this image was 1 minute.

Protoplasts were extracted in 50mM Tris, 10 mM EDTA and Triton X100, 0.02%.

immunofluorescence using anti-Arf1 antibodies

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

Additional information

Cellular [compartment marker] of Golgi in immunolocalization and COP1 in western blot

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.

Related products

AS08 327 | Anti-Sec21 (gamma subunit, COP vesicles)(Golgi marker in immunolocalization and COP1 marker in western blot), rabbit antibodies

AS08 325PRE | Arf1 | ADP-ribosylation factor 1, pre-immune serum

recommended secondary antibody

Plant and algal protein extraction buffer

Secondary antibodies

Background

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.

Product citations

Selected references Singh et al. (2018). A single class of ARF GTPase activated by several pathway-specific ARF-GEFs regulates essential membrane traffic in Arabidopsis. PLoS Genet. 2018 Nov 15;14(11):e1007795. doi: 10.1371/journal.pgen.1007795.
Gonzaga Heredia-Martinez et al. (2018). Chloroplast damage induced by the inhibition of fatty acid synthesis triggers autophagy in Chlamydomonas. Plant Physiol, Sept. 2018.
Lynch et al. (2017). Multifaceted plant responses to circumvent Phe hyperaccumulation by downregulation of flux through the shikimate pathway and by vacuolar Phe sequestration. Plant J. 2017 Dec;92(5):939-950. doi: 10.1111/tpj.13730.
Vincent et al. (2017). A genome-scale analysis of mRNAs targeting to plant mitochondria: upstream AUGs in 5' untranslated regions reduce mitochondrial association. Plant J. 2017 Dec;92(6):1132-1142. doi: 10.1111/tpj.13749.
Ma et al. (2016). Phosphatidylserine Synthase Controls Cell Elongation Especially in the Uppermost Internode in Rice by Regulation of Exocytosis. PLoS One. 2016 Apr 7;11(4):e0153119. doi: 10.1371/journal.pone.0153119. eCollection 2016.
Yüzbaşıoğlu et al. (2016). Functional specialization of Arf paralogs in nodules of model legume, Medicago truncatula. Plant Growth Regul. DOI: 10.1007/s10725-016-0227-2.
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)