AS07 268 | Clonality: Polyclonal | Host: Rabbit | Reactivity: higher plants
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0.5 µg/ml (ELISA), 1 ug/10 ml of incubation buffer (WB), 1: 40 (IL)
|Expected | apparent MW||
10 - 100 for various glycoproteins
|Confirmed reactivity||higher plants|
|Not reactive in||
no confirmed exceptions from predicted reactivity known in the moment
PLA2 (phospholipase 2 from bee venom) which contains only α1.3 fucose, Sigma, product number P9279
Type II - horseradish peroxidase which contains β1.2 Xylose and α1.3 fucose, Sigma, product number P8250
The antibody does not recognize alpha 1,6-fucose.
|Selected references||Nakanishi et al. (2017). Protection of Human Colon Cells from Shiga Toxin by Plant-based Recombinant Secretory IgA. Sci Rep. 2017 Apr 3;7:45843. doi: 10.1038/srep45843. (ELISA)
Hanania et al. (2017). Establishment of a tobacco BY2 cell line devoid of plant specific xylose and fucose as a platform for the production of biotherapeutic proteins. Plant Biotechnol J. 2017 Feb 3. doi: 10.1111/pbi.12702.
Ebert et al. (2015). Identification and Characterization of a Golgi-Localized UDP-Xylose Transporter Family from Arabidopsis. Plant Cell. 2015 Mar 24. pii: tpc.114.133827.
Lehtimäki et al. (2014). Posttranslational modifications of FERREDOXIN-NADP+ OXIDOREDUCTASE in Arabidopsis chloroplasts. Plant Physiol. 2014 Dec;166(4):1764-76. doi: 10.1104/pp.114.249094
Application example 1
Total cell extract from Arabidopsis thaliana wild type (1) and cell extracts from different mutants defective in fucosyltransferases (2-5) (data not published yet).
Primary antibody has been used at 10 µg/10 ml of incubation buffer. Detection has been done using enchanced chemiluminescence (ECL).
Application example 2
Dot blot reaction of anti-Fucose and anti-Xylose antibodies with various controls: Avidin (Fuc+/Xyl+), Fetuin (Fuc-/Xyl-), PLA2 (Fuc+/Xyl-) and Mur1-2 (Fuc-/Xyl+). 2 µl of each extract were spotted on a nitrocellulose membrane, placed on top of 2 WHATMAN filters (one soaked in TBS-T) and dried for 1.5 h at RT. The mem-brane was blocked for 30 min with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and incubated with anti-Fucose(1) (AS07 268, 1:1000) or anti-Xylose(2) (AS07 267, 1:1000) for 30 min and then with secondary anti-rabbit(1:1000) antibody (ALP conjugated, recommended secondary antibody AS09 607). Membrane was washed with TBS-T 3 x 10 minutes before reaction development using alkaline phosphatase reagent BCIP®/NTB premixed solution (Sigma, Prod. No. B6404).
Please follow this link for a more detailed Dot-Blot protocol
Application example 3
Immunolocalization using anti-fucose antibodies on a 40µm vibratome section of a 21 day old Arabidopsis thaliana hypocotyl (wood). Fixation: 4% formaldehyde fixation in 1X PME (100 mM PIPES, 1 mM MgSO4, 2 mM EGTA); blocking: 5 % BSA; primary antibody dilution: 1:40; secondary antibody: 1:300 AlexaFluor568 from Invitrogen (Life Technologies), Alexa Fluor® 568 Goat Anti-Rat IgG (H+L); Left panel, immunolabel channel (cyan) overlaid on grayscale channel (0.001% Calcofluor white which stains cellulose and hemicellulose). Right panel, only immunolabelling
Courtesy of Dr. Hardy Hall and Dr. Urs Fischer, Umeå Plant Science Centre, Sweden
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