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product information
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| background |
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Alcohol dehydrogenase (E.C.:1.1.1.1) is an important enzyme for plants and microbes. In microalgae and bacteria the conversion of Acetyl-CoA to ethanol under conditions of oxygen deprivation is catalyzed by the dual function enzyme alcohol/acetaldehyde dehydrogenase (ADH/ALDH; E.C.:1.1.1.1 /1.2.1.10). This reaction results in NAD+ recycling and allows glycolysis to proceed. |
| immunogen |
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KLH-conjugated peptide derived from available algal and bacterial ADH sequences including Chlamydomonas reinhardtii |
| antibody format |
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rabbit |
polyclonal |
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serum |
lyophilized |
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| quantity |
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200 µl |
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 |
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western blot (WB) |
| related products |
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AS10 685 | Anti-plant ADH | alcohol dehydrogenase (hypoxia marker) AS10 691 | PDC | pyruvate decarboxylase |
| additional information |
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selected peptide is well conserved in Escherichia coli ADHE, most of the microbial dual function aldehyde/alcohol dehydrogenases (ADHE) and Iron-containing alcohol dehydrogenases are also conserved in a peptide used to elicit ADH antibody. |
application information
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| recommended dilution |
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1: 1000 with standard ECL (WB) |
| expected | apparent MW |
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100 | 100 kDa (C.reinhardtii), 96 kDa (E.coli)
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| confirmed reactivity |
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Chlamydomonas reinhardtii, E.coli
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| predicted reactivity |
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algae |
| not reactive in |
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no confirmed exceptions from predicted reactivity known in the moment |
| additional information |
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to be added when available |
| selected references |
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to be added when available, antibody released in April 2011 |
application example 
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
||| For applications or usage on species others than stated as confirmed above Agrisera offers a payment-after-testing option. To learn more about this or for any questions on this product, please use the LiveChat option in the left menue bar or contact us at support@agrisera.com
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