Journal: Human Molecular Genetics

Article Title: Genetic absence of ALOX5 protects from homocysteine-induced memory impairment, tau phosphorylation and synaptic pathology

doi: 10.1093/hmg/ddx088

Figure Lengend Snippet: Diet-induced high Hcy-dependent increase in tau phosphorylation is prevented by genetic absence of ALOX5. (A) Representative western blots of soluble total tau, phosphorylated tau at residues T231/S235 (AT180), at T181 (AT270), S396/S404 (PHF1) and S396 (PHF13) in brain cortex homogenates from wild type mice receiving regular diet (WT); wild type mice receiving folate and B vitamin deficient diet (WT + Diet); 5LOKO mice receiving regular diet (5LOKO); and 5LOKO mice receiving the Diet (5LOKO + Diet). (B) Densitometric analyses of the immunoreactivities to the antibodies shown in the previous panel (*P < 0.05). Values represent mean ± S.E.M. (n = 5 WT-control, n = 5 WT + Diet, n = 5 5LOKO-control, n = 5 5LOKO + Diet).

Article Snippet: After three washings with T-TBS (pH7.4), membranes were incubated with IRDye 800CW-labeled secondary antibodies (LI-COR Bioscience, Lincoln, NE) at RT for 1 h. Signals were developed with Odyssey Infrared Imaging Systems (LI-COR Bioscience). β-Actin was always used as internal loading control. table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Immunogen Host Dilution Source Catalog number Tau-1 Bovine microtubule associated protein Mouse 1:200 Cell signaling 4019 AT-180 Peptide containing phospho-T231/S235 Mouse 1:200 Thermo P10636 AT-270 Peptide containing phospho-T181 Mouse 1:300 Thermo MN1050 PHF-13 Peptide containing phospho-Ser396 Mouse 1:300 Cell signaling 9632 PHF-1 Peptide containing phosphor-Ser396/S404 Mouse 1:200 Dr Davies P. N/A PSD95 Purified recombinant rat PSD-95 Mouse 1:200 Thermo MA1-045 SYP aa 221–313 of SYP of human origin Mouse 1:400 Santa Cruz sc-55507 MAP2 Purified microtubule-associated protein from rat brain Rabbit 1:300 Millipore AB5622 GFAP Spinal cord homogenate of bovine origin Mouse 1:300 Santa Cruz sc-33673 CD45 aa 1075–1304 of CD45 of human origin Rabbit 1:200 Santa Cruz sc-25590 Actin Gizzard Actin of avian origin Mouse 1:1000 Santa Cruz sc-47778 Open in a separate window Antibodies used in the study

Techniques: Western Blot

Journal: Human Molecular Genetics

Article Title: Genetic absence of ALOX5 protects from homocysteine-induced memory impairment, tau phosphorylation and synaptic pathology

doi: 10.1093/hmg/ddx088

Figure Lengend Snippet: Antibodies used in the study

Article Snippet: After three washings with T-TBS (pH7.4), membranes were incubated with IRDye 800CW-labeled secondary antibodies (LI-COR Bioscience, Lincoln, NE) at RT for 1 h. Signals were developed with Odyssey Infrared Imaging Systems (LI-COR Bioscience). β-Actin was always used as internal loading control. table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Immunogen Host Dilution Source Catalog number Tau-1 Bovine microtubule associated protein Mouse 1:200 Cell signaling 4019 AT-180 Peptide containing phospho-T231/S235 Mouse 1:200 Thermo P10636 AT-270 Peptide containing phospho-T181 Mouse 1:300 Thermo MN1050 PHF-13 Peptide containing phospho-Ser396 Mouse 1:300 Cell signaling 9632 PHF-1 Peptide containing phosphor-Ser396/S404 Mouse 1:200 Dr Davies P. N/A PSD95 Purified recombinant rat PSD-95 Mouse 1:200 Thermo MA1-045 SYP aa 221–313 of SYP of human origin Mouse 1:400 Santa Cruz sc-55507 MAP2 Purified microtubule-associated protein from rat brain Rabbit 1:300 Millipore AB5622 GFAP Spinal cord homogenate of bovine origin Mouse 1:300 Santa Cruz sc-33673 CD45 aa 1075–1304 of CD45 of human origin Rabbit 1:200 Santa Cruz sc-25590 Actin Gizzard Actin of avian origin Mouse 1:1000 Santa Cruz sc-47778 Open in a separate window Antibodies used in the study

Techniques: Purification, Recombinant