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Plant/Algal cell antibodies / Chlamydomonas reinhardtii


Arf1 | ADP-ribosylation factor 1

Art no: AS08 325
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product information

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.

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.

tested applications  

western blot (WB), immunofluorescence (IF)

related products  

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

recommended secondary antibody

additional information  

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

application information

recommended dilution  

1:1000 with standard ECL (WB), 1: 1000 (IF)

expected | apparent MW  

21 kDa (Arabidopsis thaliana)

confirmed reactivity  

Arabidopsis thaliana, Lilium longiflorum, Nicotiana tabacum, Chlamydomonas reinhardtii, moss: Physcomitrella patens

predicted reactivity  

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

not reactive in  

Microsporidia sp.

additional information  

to be added when available 

selected references  

Wulfetange et al. (2011). The Cytokiin Receptors of Arabidopsis thaliana are Locating Mainly to the Endoplasmic Reticulum. Plant Physiol. (in press).

Lang, E.G.E., S.J. Mueller, S.N.W. Hoernstein, J. Porankiewicz-Asplund, M. Vervliet-Scheebaum, R. Reski (2010). Simultaneous isolation of pure and intact chloroplasts and mitochondria from moss as basis for sub-cellular proteomics. Plant Cell Reports, DOI: 10.1007/s00299-010-0935-4. (open source)

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 (recommended secondary antibody AS09 602). Signal was detected with standard ECL and exposure time for this image was 1 minute.

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

Western blot detection using Arf1 antibody

 

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

immunofluorescence using anti-Arf1 antibodies


||| For applications or usage on species others than stated above Agrisera offers a payment-after-testing option. To learn more about this or for any questions on this product, please use LiveChat option in the left menu or contact us at support@agrisera.com

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