ADH/ALDH | Alcohol/acetaldehyde dehydrogenase (bacterial/algal)
AS10 748 | clonality: polyclonal | host: rabbit | reactivity: C. reinhardtii, E.coli
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|Recommended dilution||1 : 1000 (WB)|
|Expected | apparent MW||
100 | 100 kDa (C.reinhardtii), 96 kDa (E.coli)
|Confirmed reactivity||Chlamydomonas reinhardtii, E.coli, Streptococcus pmenumoniae
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
|Selected references||Laurenceau et al. (2015). Conserved Streptococcus pneumoniae Spirosomes Suggest a Single Type of Transformation Pilus in Competence. PLoS Pathog. 2015 Apr 15;11(4):e1004835. doi: 10.1371/journal.ppat.1004835.
Kukuczka et al. (2014). Proton Gradient Regulation5-Like1-Mediated Cyclic Electron Flow Is Crucial for Acclimation to Anoxia and Complementary to Nonphotochemical Quenching in Stress Adaptation. Plant Physiol. 2014 Jun 19;165(4):1604-1617.
30 μg of a total cell extract from Chlamydomonas reinhardtii and E.coli strains DC272 and DC271 were loaded on Criterion™,Tris-HCl 10% polyacrylamide gels (Biorad) and molecular weight compared to those of the PageRuler™ Plus Prestained Protein Ladder (Fermentas). After SDS-PAGE, gels were transferred to PVDF membranes (Biorad) by the Trans-Blot SD semidry Transfer Cell method (Biorad) for 1 hour at 10V. Blocking of the PVDF membrane has been done for 3 hours in TBST milk 5% and has been followed by overnight incubation at 4°C with the primary anti-ADH/ALDH antibodies 1:1000 in TBST milk 1%. After three intensive washes, the membrane was incubated for one hour at room temperature with the secondary HRP-conjugated goat anti-rabbit (Agrisera AS09 602, in 1:50 000 dilution in TBST milk 1%). After three washes with TBST (10 minutes each), detection was achieved by the Amersham ECL™ Western Blotting System. Exposure time was 2 minutes for Chlamydomonas reinhardtii sample and 10 seconds for E. coli samples.
E. coli strains DC272 and DC271 were provided by Professor David P. Clark, Souther Illinois University. The DC272 mutant strain is misregulated in AdhE expression so that the bacteria expresses the ADHE protein constitutively.
Courtesy Dr. Leonardo Magneschi, Scuola Superiore Sant'Anna, Italy
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