rabbit scad  (Proteintech)

Journal: iScience

Article Title: Ginsenoside Rb1 regulates CPT1A deacetylation to inhibit intramuscular fat infiltration after rotator cuff tear

doi: 10.1016/j.isci.2024.110331

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-ACADS , Proteintech , Cat# 16623-1-AP; RRID: AB_10666165.

Techniques: Recombinant, Modification, Staining, H&E Stain, Mutagenesis, Software

Journal: iScience

Article Title: Mitochondrial glycerol 3-phosphate dehydrogenase deficiency exacerbates lipotoxic cardiomyopathy

doi: 10.1016/j.isci.2024.109796

Figure Lengend Snippet: Downregulated mGPDH/SIRT5 signaling inhibits the desuccinylation of FAO proteins and promotes ACADS degradation (A) Schematic of experimental workflow for the identification of lysine succinylation substrates by proteomics in heart lysates. (B) The distribution of the number of lysine succinylation site identifications per protein was shown. (C) Top 10 terms of KEGG pathway analysis in succinylated protein targeted by mGPDH cKO under HFD condition. (D) Schematic represents 4 steps and 15 enzymes (including 4 auxiliary enzymes) of mitochondrial FAO. 14 of them displayed an altered succinylation level after mGPDH cKO, 13 were increased (highlighted with red color) and 1 was decreased (blue). The rest one which succinylation level could not be detected was shown in white. These enzymes are showing by their gene name. (E) Heatmap shown the changes in the levels of lysine succinylation on FAO enzymes according to our quantitative proteomics. (F and G) Heart lysates from HFD-fed mGPDH cKO and Ctrl mice (F), and cell lysates from H9C2 cells with mGPDH specific siRNA transfected and PA-treated (G) were immunoprecipitated with antibodies against ACADS or ACADM, and succinylation level of these two proteins and its quantifications were shown. (H) PA-treated H9C2 cells were transfected with siRNA for mGPDH and overexpressing plasmids for SIRT5_WT or catalytically inactive SIRT5-H158Y mutant, cell lysates were immunoprecipitated with antibodies against ACADS, and succinylation level of ACADS and its quantifications were assessed. (I) PA-treated H9C2 cells were either untreated or treated with the proteasome inhibitors MG132 (MG), ACADS levels were analyzed by western blot and its quantifications were shown. (J) PA-treated H9C2 cells were transfected with siRNA for mGPDH, cell lysates were immunoprecipitated with antibodies against ACADS, and ubiquitinated ACADS levels were assessed. (K–M) PA-treated H9C2 cells were transfected with siRNA for mGPDH and overexpressing plasmids for HA-tagged ACADS_WT, ACADS-K250R and ACADS-K250E, succinylation level of ACADS (K), ACADS protein (L) and ubiquitination of ACADS (M) were assessed. Quantifications of K and L were shown. n = 3 mice per group for B-E, n = 6 mice per group for F, n = 3 per group for G-M. The data are presented as the means ± S.E.M. ∗∗∗ p < 0.001 by two-tailed unpaired Student’s t test.

Article Snippet: Rabbit polyclonal anti-ACADS antibody , Proteintech , Cat# 16623-1-AP; RRID: AB_10666165.

Techniques: Quantitative Proteomics, Transfection, Immunoprecipitation, Mutagenesis, Western Blot, Ubiquitin Proteomics, Two Tailed Test

Journal: iScience

Article Title: Mitochondrial glycerol 3-phosphate dehydrogenase deficiency exacerbates lipotoxic cardiomyopathy

doi: 10.1016/j.isci.2024.109796

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-ACADS antibody , Proteintech , Cat# 16623-1-AP; RRID: AB_10666165.

Techniques: Virus, Recombinant, Modification, Transfection, Labeling, XF Assay, Isolation, Staining, Bicinchoninic Acid Protein Assay, Immunoprecipitation, Plasmid Preparation, Control, Software

Journal: Current Research in Physiology

Article Title: Increased fatty acid oxidation enzyme activity in the hearts of mice fed a high fat diet does not correlate with improved cardiac contractile function

doi: 10.1016/j.crphys.2020.11.001

Figure Lengend Snippet: Loss of GCN5L1 expression prevents HFD-induced fatty acid oxidation enzyme hyperacetylation. The acetylation status of ( A, B ) LCAD and ( C, D ) SCAD was significantly increased in the hearts of WT mice, but not GCN5L1 KO mice, after a 24 week HFD. N ​= ​4–5, ∗ ​= ​ P < 0.05 vs. WT LFD.

Article Snippet: Protein expression was analyzed using the following primary antibodies: rabbit acetyl-lysine (Ac–K, #9441) from Cell Signaling Technology; rabbit long-chain acyl-CoA dehydrogenase (LCAD, #17526-1-AP) and rabbit short-chain acyl-CoA dehydrogenase (SCAD, #16623-1-AP) from Proteintech.

Techniques: Expressing

Journal: Current Research in Physiology

Article Title: Increased fatty acid oxidation enzyme activity in the hearts of mice fed a high fat diet does not correlate with improved cardiac contractile function

doi: 10.1016/j.crphys.2020.11.001

Figure Lengend Snippet: Fatty acid oxidation activity is increased in isolated cardiac lysates from HFD-fed WT mice. The enzymatic activity of ( A ) LCAD and ( B ) SCAD was significantly increased in cardiac lysates from WT mice, but not GCN5L1 KO mice, after a 24 week HFD. N ​= ​5–10, ∗ ​= ​ P < 0.05 vs. WT LFD.

Article Snippet: Protein expression was analyzed using the following primary antibodies: rabbit acetyl-lysine (Ac–K, #9441) from Cell Signaling Technology; rabbit long-chain acyl-CoA dehydrogenase (LCAD, #17526-1-AP) and rabbit short-chain acyl-CoA dehydrogenase (SCAD, #16623-1-AP) from Proteintech.

Techniques: Activity Assay, Isolation

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Acetylation of mitochondrial proteins by GCN5L1 promotes enhanced fatty acid oxidation in the heart

doi: 10.1152/ajpheart.00752.2016

Figure Lengend Snippet: Expression of cardiac fuel metabolism genes. High-fat diet (HFD) promotes an enhanced fatty acid oxidation phenotype in the heart. A and B: no significant difference was observed in short-chain acyl-CoA dehydrogenase (Acads, SCAD) or medium-chain acyl-CoA dehydrogenase (Acadm, MCAD) levels. C–G: significant increases in the mRNA content of long-chain acyl-CoA dehydrogenase (Acadl, LCAD), carnitine palmitoyltransferase 1b (Cpt1b), Cd36, pyruvate dehydrogenase kinase 4 (Pdk4), and peroxisome proliferator-activted receptor-α (Ppara) were observed with HFD feeding, whereas peroxisome proliferator-activated receptor-γ coactivator-1α (Ppargc1a; H) was significantly decreased. Values are expressed as means ± SE; n = 4 per group. P value significance is as shown in the graphs using a two-way Student’s t-test.

Article Snippet: Protein expression was analyzed using the following primary antibodies: rabbit SIRT3, rabbit acetyl-lysine (Ac-K), rabbit glutamate dehydrogenase (GDH), rabbit PDH, and rabbit cytochrome c oxidase subunit 4 (COX IV) from Cell Signaling Technologies; rabbit phospho-PDH (Ser 293 ) from Novus; rabbit LCAD, rabbit short-chain acyl-CoA dehydrogenase (SCAD), and rabbit hydroxyacyl-CoA dehydrogenase (HADHA) from Proteintech; and GCN5L1 as previously reported ( 35 ).

Techniques: Expressing

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Acetylation of mitochondrial proteins by GCN5L1 promotes enhanced fatty acid oxidation in the heart

doi: 10.1152/ajpheart.00752.2016

Figure Lengend Snippet: Impact of HFD on metabolic enzyme acetylation status. HFD feeding led to increased fatty acid oxidation (FAO) enzyme acetylation, which contributes to upregulated fatty acid utilization in diet-induced obese mice. A–C: the acetylated lysine pulldown showed an increase in the acetylation levels of the FAO proteins SCAD, LCAD, and hydroxyacyl-CoA dehydrogenase (HADHA) and the pyruvate oxidation enzyme pyruvate dehydrogenase (PDH; D) from HFD-fed cardiac tissue relative to chow-fed controls. Values are expressed as means ± SE; n = 4 per group. P value significance is as shown in the graphs using two-way Student’s t-test.

Article Snippet: Protein expression was analyzed using the following primary antibodies: rabbit SIRT3, rabbit acetyl-lysine (Ac-K), rabbit glutamate dehydrogenase (GDH), rabbit PDH, and rabbit cytochrome c oxidase subunit 4 (COX IV) from Cell Signaling Technologies; rabbit phospho-PDH (Ser 293 ) from Novus; rabbit LCAD, rabbit short-chain acyl-CoA dehydrogenase (SCAD), and rabbit hydroxyacyl-CoA dehydrogenase (HADHA) from Proteintech; and GCN5L1 as previously reported ( 35 ).

Techniques:

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Acetylation of mitochondrial proteins by GCN5L1 promotes enhanced fatty acid oxidation in the heart

doi: 10.1152/ajpheart.00752.2016

Figure Lengend Snippet: Impact of HFD-related acetylation on in vitro FAO enzyme activity. A and B: enzymatic activity of SCAD and LCAD was significantly elevated in cardiac tissues from HFD-fed mice. C: regression analysis showed a significant correlation between LCAD acetylation status and enzymatic activity from all tested mice. Values are expressed as means ± SE, n = 4 per group. P value significance is as shown in the graphs using two-way Student’s t-test.

Article Snippet: Protein expression was analyzed using the following primary antibodies: rabbit SIRT3, rabbit acetyl-lysine (Ac-K), rabbit glutamate dehydrogenase (GDH), rabbit PDH, and rabbit cytochrome c oxidase subunit 4 (COX IV) from Cell Signaling Technologies; rabbit phospho-PDH (Ser 293 ) from Novus; rabbit LCAD, rabbit short-chain acyl-CoA dehydrogenase (SCAD), and rabbit hydroxyacyl-CoA dehydrogenase (HADHA) from Proteintech; and GCN5L1 as previously reported ( 35 ).

Techniques: In Vitro, Activity Assay

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Acetylation of mitochondrial proteins by GCN5L1 promotes enhanced fatty acid oxidation in the heart

doi: 10.1152/ajpheart.00752.2016

Figure Lengend Snippet: Impact of GCN5L1 knockdown on cardiac FAO enzyme acetylation and activity. GCN5L1 promoted the acetylation and activity of cardiac FAO proteins. A and B: our immunoprecipitation-based acetylation assay showed a large decrease in the acetylation status of SCAD and LCAD proteins in GCN5L1 knockdown cells relative to the control. C and D: this decrease correlated with reduced enzymatic activities of SCAD and LCAD in vitro. Values are expressed as means ± SE; n = 4 per group. P value significance is as shown in the graphs using two-way Student’s t-test.

Article Snippet: Protein expression was analyzed using the following primary antibodies: rabbit SIRT3, rabbit acetyl-lysine (Ac-K), rabbit glutamate dehydrogenase (GDH), rabbit PDH, and rabbit cytochrome c oxidase subunit 4 (COX IV) from Cell Signaling Technologies; rabbit phospho-PDH (Ser 293 ) from Novus; rabbit LCAD, rabbit short-chain acyl-CoA dehydrogenase (SCAD), and rabbit hydroxyacyl-CoA dehydrogenase (HADHA) from Proteintech; and GCN5L1 as previously reported ( 35 ).

Techniques: Knockdown, Activity Assay, Immunoprecipitation, Acetylation Assay, Control, In Vitro