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product information
|
| background |
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DHAR1 ( Dehydroascorbate Reductase 1) the protein is induced by jasmonic acid and oxidative chemical stresses and is a key component of the ascorbate recycling system. Involved in redox homeostasis under biotic and abiotic inducers. Localized in mitochondria. Synonymes: glutathione-dependent dehydroascorbate reductase 1, chloride intracellular channel homolog 1, CLIC homolog 1, glutathione-dependent dehydroascorbate reductase 1, AtDHAR1, GSH-dependent dehydroascorbate reductase 1, mtDHAR, AT1G19570. |
| immunogen |
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KLH-conjugated synthetic peptide derived from known DHAR1 sequence of Arabidopsis thaliana Q9FWR4 |
| antibody format |
|
rabbit |
polyclonal |
|
affinity purified serum |
lyophilized |
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| quantity |
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200 µg |
for reconstitution add 200 µl, of sterile water. |
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| storage |
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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) |
| related products |
|
AS11 1747 | Anti-DHAR2, rabbit antibodies |
| additional information |
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to be added when available |
application information
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| recommended dilution |
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1 : 5000 with standard ECL (WB) |
| expected | apparent MW |
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23.6 | 23.4 kDa |
| confirmed reactivity |
|
Arabidopsis thaliana |
| predicted reactivity |
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dicots including: Nicotiana tabacum, Solanum lycopersicum, Solanum tuberosum, Ricinus communis, Zinnia elegans, monocots including: Triticum aestivum, trees: Populus trichocarpa |
| not reactive in |
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no confirmed exceptions from predicted reactivity are currently known |
| additional information |
|
to be added when available |
| selected references |
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Grefen et al. (2009). The determination of protein-protein interactions by the mating-based split-ubiquitin system (mbSUS). Methods Mol Biol 479:217-233. |
application example 
1cm2 of a leaf from Arabidopsis thaliana Col-0 (1) and or t-DNA insertion lines dhar1-1 (2), dhar1-2 (3), dhar1-3 (4), dhar2-1 (5), dhar2-2 (6), dhar1-3 EOS-DHAR1 (7), was extracted using 200µl Lyse&Load-Buffer (Grefen et al. 2009). 10 µl were separated on a 15% SDS-PAGE and blotted 1h to PVDF (using Bjerrum Buffer in a semidry blot). Blots were blocked with 5% Milk in 1xTBS-Tween20 (1%) for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1:5000 (in 5% Milk 1xTBS-Tween20 (1%) + 0.01 % NaN3) ON at 4°C with agitation. The antibody solution was decanted and the blot was washed 3 times for 10 minutes with 1x TBS-Tween20 at RT with agitation. Blot was incubated in secondary antibody BioRad anti-rabbit IgG AP-conjugate (#170-6518) diluted to 1:2000 in 5% Milk 1xTBS-Tween20 (1%) + 0.01 % NaN3 for 1h at RT with agitation. The blot was washed as above, equilibrated in staining buffer (100mM Tris-HCl, 100mM NaCl, 5mM MgCl2, see Grefen et al. 2009) and developed for 5-15 min. with staining solution (Nitro blue tetrazolium chloride (NBT) and 5-bromo-4-chloro-3-indoylphosphate-p-toluidin (BCIP) in staining buffer). Courtesy Dr. Chrisopher Grefen, UK
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