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ADH/ALDH | Alcohol/acetaldehyde dehydrogenase (bacterial/algal)

AS10 748  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: C. reinhardtii, E.coli

ADH/ALDH | Alcohol/acetaldehyde dehydrogenase (bacterial/algal) in the group Antibodies Plant/Algal  / Environmental Stress / Hypoxia at Agrisera AB (Antibodies for research) (AS10 748)
ADH/ALDH | Alcohol/acetaldehyde dehydrogenase (bacterial/algal)



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Product Information

Immunogen

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

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 200 ĩl
Reconstitution For reconstitution add 200 ĩl of sterile water
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications Western blot (WB)
Recommended dilution 1 : 1000 (WB)
Expected | apparent MW

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

Reactivity

Confirmed reactivity Chlamydomonas reinhardtii, E.coli, Streptococcus pmenumoniae
Predicted reactivity Algae, Nannochloropsis gaditana
Species of your interest not listed? Contact us
Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples

Application example

western blot detection using anti-bacterial ADH antibodies

 

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 using chemiluminescent detection reagent. 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
 

Additional information

Additional information Selected peptide is well conserved in Escherichia coli ADHE (P0A9Q7), 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

Related products

Background

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.

Product citations

Selected references Kurylo et al. (2018). Endogenous rRNA Sequence Variation Can Regulate Stress Response Gene Expression and Phenotype. Cell Rep. 2018 Oct 2;25(1):236-248.e6. doi: 10.1016/j.celrep.2018.08.093.
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.
immunogen:

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

Reconstitution: For reconstitution add 200 ĩl of sterile water
Host: Rabbit
Clonality: Polyclonal
Purity: Serum
Format: Lyophilized
Quantity: 200 ĩl
storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications: Western blot (WB)
recommended dilution: 1 : 1000 (WB)
calculated | apparent molecular mass [kDa]:

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

Confirmed reactivity: Chlamydomonas reinhardtii, E.coli, Streptococcus pmenumoniae
predicted reactivity: Algae, Nannochloropsis gaditana
Species of your interest not listed? Contact us
not reactive in: No confirmed exceptions from predicted reactivity are currently known
Picture (footer):

Application example

western blot detection using anti-bacterial ADH antibodies

 

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 using chemiluminescent detection reagent. 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
 

additional information: Selected peptide is well conserved in Escherichia coli ADHE (P0A9Q7), 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
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.

All references: Kurylo et al. (2018). Endogenous rRNA Sequence Variation Can Regulate Stress Response Gene Expression and Phenotype. Cell Rep. 2018 Oct 2;25(1):236-248.e6. doi: 10.1016/j.celrep.2018.08.093.
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.

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