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Plant/Algal cell antibodies
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HSP70 | heat shock protein 70, mitochondrial
AS08 347 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana
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| application information |
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| recommended dilution | 1: 4000 (WB) |
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| expected | apparent MW | 73 | 70 kDa |
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| confirmed reactivity | Arabidopsis thaliana |
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| predicted reactivity | dicots including: Medicago truncatula, Spinacia oleracea, Phaseolus vulgaris, Pisum sativum, Solanum tuberosum, Vitis vinifera, monoctos including: Oryza sativa, Triticum aestivum, Zea mays, trees: Populus trichocarpa |
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| not reactive in | no confirmed exceptions from predicted reactivity known in the moment |
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| additional information | to be added when available |
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| selected references | to be added when available, antibody released in October 2009 |
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application example 25 μg of Arabidopsis thaliana leaf extract and 15 μg)Arabidopsis thaliana mitochondrial fraction were separated on 10% gel and blotted on nitrocellulose membrane using wet transfer (0.22% CAPS, pH 11). Filters where blocked (1.5h) in 5% milk in TBST (1X TBS, 0,1% Tween 20), incubated with 1: 1000 anti-HSP70 mitochondrial antibodies (2h in TBST) followed by incubation with 1: 10 000 secondary anti-rabbit (1h) HRP-coupled antibodies and visualized with standard ECL on Kodak autoradiography film for 15-60 s. Mitochondria were isolated as described by Urantowka et al. (Plant Mol Biol, 2005, 59:239-52). Mitochondrial pellets were suspended in 1X Laemmli buffer (5% beta-mercaptoetanol, 3.7% glycerol, 1.1% SDS, 23 mM Tris- HCl pH 6.8, 0.01% bromophenol blue), heated (95°C, 5 min.) and centrifuged (13 000rpm, 1 min.). Leaf extracts were prepared as described by Martinez-Garcia et al. (Plant J., 1999, 20:251-7). Courtesy Dr. Janusz Piechota, Wrocław University, Poland
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