product information

background
 

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
 

KLH-conjugated peptide derived from available algal and bacterial ADH sequences including Chlamydomonas reinhardtii

antibody format
 

rabbit

polyclonal

serum

lyophilized

quantity
 

200 µl

for reconstitution add 200 µl of sterile water

storage
 

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
 

AS10 685 | Anti-plant ADH | alcohol dehydrogenase (hypoxia marker)

AS10 691 | PDC | pyruvate decarboxylase

additional information
 

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

recommended dilution
 

1: 1000 with standard ECL (WB)

expected | apparent MW
 

100 | 100 kDa (C.reinhardtii), 96 kDa (E.coli)

confirmed reactivity
 

Chlamydomonas reinhardtii, E.coli

predicted reactivity
 

algae

not reactive in
 

no confirmed exceptions from predicted reactivity known in the moment

additional information
 

to be added when available

selected references
 

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
 

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