rabbit polyclonal anti-nd6  (Absolute Biotech Inc)

Journal: Scientific Reports

Article Title: Evaluation of mitochondrial bioenergetics, dynamics, endoplasmic reticulum-mitochondria crosstalk, and reactive oxygen species in fibroblasts from patients with complex I deficiency

doi: 10.1038/s41598-018-19543-3

Figure Lengend Snippet: ND6 (complex I subunit) ( A ), NDUFV1 (complex I subunit) ( B ), ACAD9 (complex I assembly factor) ( C ), SDHB (complex II subunit), UQCR2 (complex III subunit) and MTCO1 (complex IV subunit) ( D ) protein content in mitochondria prepared from ND6 deficient, NDUFV1 deficient and ACAD9 deficient fibroblasts cultured in media with or without glucose for 48 h. GAPDH was used as loading control ( A – C , and E ).

Article Snippet: Following electrophoresis, the gel was blotted onto a nitrocellulose membrane, which was incubated with rabbit anti-ND6 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-NDUFV1 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-ACAD9 antiserum (1:500), Cocalico Biologicals Inc., PA, rodent anti-total OXPHOS cocktail antibody (1:250), Abcam, Cambridge, MA, mouse anti-mitofusin 1 (MFN1) monoclonal antibody (1:100), Abcam, Cambridge, MA, mouse anti-dynamin-related protein 1 (DRP1) monoclonal antibody (1:100), Abcam, Cambridge, MA, rabbit anti-very long-chain acyl-CoA dehydrogenase (VLCAD) antiserum (1:1,000), Cocalico Biologicals Inc., PA, rabbit anti-voltage-dependent anion channel 1 (VDAC1) monoclonal antibody (1:1,000), Abcam, Cambridge, MA, mouse anti-glucose-related protein 75 (Grp75) monoclonal antibody (1:250), Abcam, Cambridge, MA, rabbit anti-glucose-related protein 78 (Grp78) polyclonal antibody (1:250), Abcam, Cambridge, MA, mouse anti-DNA damage inducible transcript 3 (DDIT3) monoclonal antibody (1:250), Abcam, Cambridge, MA, goat anti-inositol 1,4,5-trisphosphate receptor 3 (IP3R) polyclonal antibody (1:50), Santa Cruz Biotechnology, Dallas, TX, or IgG-HRP conjugated antibody, Bio-Rad, Hercules, CA.

Techniques: Cell Culture, Control

Journal: Scientific Reports

Article Title: Evaluation of mitochondrial bioenergetics, dynamics, endoplasmic reticulum-mitochondria crosstalk, and reactive oxygen species in fibroblasts from patients with complex I deficiency

doi: 10.1038/s41598-018-19543-3

Figure Lengend Snippet: Immunofluorescence analysis of ND6 deficient ( A ), NDUFV1 deficient ( B ) and ACAD9 deficient ( C ) fibroblasts cultured in media with or without glucose for 48 h. The merged image shows colocalization of ND6, NDUFV1 or ACAD9 visualized with green fluorescently tagged antibody and mitochondria visualized with MitoTracker Red as yellow. Nuclei were visualized with DAPI staining.

Article Snippet: Following electrophoresis, the gel was blotted onto a nitrocellulose membrane, which was incubated with rabbit anti-ND6 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-NDUFV1 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-ACAD9 antiserum (1:500), Cocalico Biologicals Inc., PA, rodent anti-total OXPHOS cocktail antibody (1:250), Abcam, Cambridge, MA, mouse anti-mitofusin 1 (MFN1) monoclonal antibody (1:100), Abcam, Cambridge, MA, mouse anti-dynamin-related protein 1 (DRP1) monoclonal antibody (1:100), Abcam, Cambridge, MA, rabbit anti-very long-chain acyl-CoA dehydrogenase (VLCAD) antiserum (1:1,000), Cocalico Biologicals Inc., PA, rabbit anti-voltage-dependent anion channel 1 (VDAC1) monoclonal antibody (1:1,000), Abcam, Cambridge, MA, mouse anti-glucose-related protein 75 (Grp75) monoclonal antibody (1:250), Abcam, Cambridge, MA, rabbit anti-glucose-related protein 78 (Grp78) polyclonal antibody (1:250), Abcam, Cambridge, MA, mouse anti-DNA damage inducible transcript 3 (DDIT3) monoclonal antibody (1:250), Abcam, Cambridge, MA, goat anti-inositol 1,4,5-trisphosphate receptor 3 (IP3R) polyclonal antibody (1:50), Santa Cruz Biotechnology, Dallas, TX, or IgG-HRP conjugated antibody, Bio-Rad, Hercules, CA.

Techniques: Immunofluorescence, Cell Culture, Staining

Journal: Scientific Reports

Article Title: Evaluation of mitochondrial bioenergetics, dynamics, endoplasmic reticulum-mitochondria crosstalk, and reactive oxygen species in fibroblasts from patients with complex I deficiency

doi: 10.1038/s41598-018-19543-3

Figure Lengend Snippet: Basal and maximal respiration of three wild type (WT) fibroblast lines ( A ). Basal respiration of ND6 deficient, NDUFV1 deficient and ACAD9 deficient fibroblasts cultured in media with ( B and D ) or without glucose for 72 h ( C and E ). Data are means ± SD. ****P < 0.0001, compared to wild type (WT) cells ( t test for unpaired samples) ( B – E ).

Article Snippet: Following electrophoresis, the gel was blotted onto a nitrocellulose membrane, which was incubated with rabbit anti-ND6 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-NDUFV1 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-ACAD9 antiserum (1:500), Cocalico Biologicals Inc., PA, rodent anti-total OXPHOS cocktail antibody (1:250), Abcam, Cambridge, MA, mouse anti-mitofusin 1 (MFN1) monoclonal antibody (1:100), Abcam, Cambridge, MA, mouse anti-dynamin-related protein 1 (DRP1) monoclonal antibody (1:100), Abcam, Cambridge, MA, rabbit anti-very long-chain acyl-CoA dehydrogenase (VLCAD) antiserum (1:1,000), Cocalico Biologicals Inc., PA, rabbit anti-voltage-dependent anion channel 1 (VDAC1) monoclonal antibody (1:1,000), Abcam, Cambridge, MA, mouse anti-glucose-related protein 75 (Grp75) monoclonal antibody (1:250), Abcam, Cambridge, MA, rabbit anti-glucose-related protein 78 (Grp78) polyclonal antibody (1:250), Abcam, Cambridge, MA, mouse anti-DNA damage inducible transcript 3 (DDIT3) monoclonal antibody (1:250), Abcam, Cambridge, MA, goat anti-inositol 1,4,5-trisphosphate receptor 3 (IP3R) polyclonal antibody (1:50), Santa Cruz Biotechnology, Dallas, TX, or IgG-HRP conjugated antibody, Bio-Rad, Hercules, CA.

Techniques: Cell Culture

Journal: Scientific Reports

Article Title: Evaluation of mitochondrial bioenergetics, dynamics, endoplasmic reticulum-mitochondria crosstalk, and reactive oxygen species in fibroblasts from patients with complex I deficiency

doi: 10.1038/s41598-018-19543-3

Figure Lengend Snippet: Mitochondrial membrane potential (ΔΨ) ( A and B ) and ATP production ( C ) in ND6 deficient, NDUFV1 deficient and ACAD9 deficient fibroblasts cultured in media with or without glucose for 72 h. Data are means ± SD. **P < 0.01, ***P < 0.001, ****P < 0.0001, compared to wild type (WT) cells ( t test for unpaired samples).

Article Snippet: Following electrophoresis, the gel was blotted onto a nitrocellulose membrane, which was incubated with rabbit anti-ND6 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-NDUFV1 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-ACAD9 antiserum (1:500), Cocalico Biologicals Inc., PA, rodent anti-total OXPHOS cocktail antibody (1:250), Abcam, Cambridge, MA, mouse anti-mitofusin 1 (MFN1) monoclonal antibody (1:100), Abcam, Cambridge, MA, mouse anti-dynamin-related protein 1 (DRP1) monoclonal antibody (1:100), Abcam, Cambridge, MA, rabbit anti-very long-chain acyl-CoA dehydrogenase (VLCAD) antiserum (1:1,000), Cocalico Biologicals Inc., PA, rabbit anti-voltage-dependent anion channel 1 (VDAC1) monoclonal antibody (1:1,000), Abcam, Cambridge, MA, mouse anti-glucose-related protein 75 (Grp75) monoclonal antibody (1:250), Abcam, Cambridge, MA, rabbit anti-glucose-related protein 78 (Grp78) polyclonal antibody (1:250), Abcam, Cambridge, MA, mouse anti-DNA damage inducible transcript 3 (DDIT3) monoclonal antibody (1:250), Abcam, Cambridge, MA, goat anti-inositol 1,4,5-trisphosphate receptor 3 (IP3R) polyclonal antibody (1:50), Santa Cruz Biotechnology, Dallas, TX, or IgG-HRP conjugated antibody, Bio-Rad, Hercules, CA.

Techniques: Membrane, Cell Culture

Journal: Scientific Reports

Article Title: Evaluation of mitochondrial bioenergetics, dynamics, endoplasmic reticulum-mitochondria crosstalk, and reactive oxygen species in fibroblasts from patients with complex I deficiency

doi: 10.1038/s41598-018-19543-3

Figure Lengend Snippet: Fatty acid oxidation (FAO) flux ( A ) and very long-chain acyl-CoA dehydrogenase (VLCAD) protein content ( B ) in ND6 deficient, NDUFV1 deficient and ACAD9 deficient fibroblasts cultured in media with or without glucose for 48 h. FAO flux was measured in fibroblasts cultured in a 6-well plate. VLCAD content was measured in mitochondria prepared from fibroblasts. Data are means ± SD. *P < 0.05, compared to wild type (WT) cells ( t test for unpaired samples).

Article Snippet: Following electrophoresis, the gel was blotted onto a nitrocellulose membrane, which was incubated with rabbit anti-ND6 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-NDUFV1 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-ACAD9 antiserum (1:500), Cocalico Biologicals Inc., PA, rodent anti-total OXPHOS cocktail antibody (1:250), Abcam, Cambridge, MA, mouse anti-mitofusin 1 (MFN1) monoclonal antibody (1:100), Abcam, Cambridge, MA, mouse anti-dynamin-related protein 1 (DRP1) monoclonal antibody (1:100), Abcam, Cambridge, MA, rabbit anti-very long-chain acyl-CoA dehydrogenase (VLCAD) antiserum (1:1,000), Cocalico Biologicals Inc., PA, rabbit anti-voltage-dependent anion channel 1 (VDAC1) monoclonal antibody (1:1,000), Abcam, Cambridge, MA, mouse anti-glucose-related protein 75 (Grp75) monoclonal antibody (1:250), Abcam, Cambridge, MA, rabbit anti-glucose-related protein 78 (Grp78) polyclonal antibody (1:250), Abcam, Cambridge, MA, mouse anti-DNA damage inducible transcript 3 (DDIT3) monoclonal antibody (1:250), Abcam, Cambridge, MA, goat anti-inositol 1,4,5-trisphosphate receptor 3 (IP3R) polyclonal antibody (1:50), Santa Cruz Biotechnology, Dallas, TX, or IgG-HRP conjugated antibody, Bio-Rad, Hercules, CA.

Techniques: Cell Culture

Journal: Scientific Reports

Article Title: Evaluation of mitochondrial bioenergetics, dynamics, endoplasmic reticulum-mitochondria crosstalk, and reactive oxygen species in fibroblasts from patients with complex I deficiency

doi: 10.1038/s41598-018-19543-3

Figure Lengend Snippet: Mitochondrial mass ( A and B ) and dynamics ( C ) in ND6 deficient, NDUFV1 deficient and ACAD9 deficient fibroblasts cultured in media with or without glucose for 48 h. For mitochondrial mass, ND6 deficient, NDUFV1 deficient and ACAD9 deficient fibroblasts were incubated with MitoTracker Green. Data are means ± SD. *P < 0.05, compared to wild type (WT) ( t test for unpaired samples) ( A and B ). For mitochondrial dynamics, dynamin-related protein 1 (DRP1) and mitofusin 1 (MFN1) protein content was evaluated in mitochondria prepared from ND6 deficient, NDUFV1 deficient and ACAD9 deficient fibroblasts ( C ).

Article Snippet: Following electrophoresis, the gel was blotted onto a nitrocellulose membrane, which was incubated with rabbit anti-ND6 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-NDUFV1 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-ACAD9 antiserum (1:500), Cocalico Biologicals Inc., PA, rodent anti-total OXPHOS cocktail antibody (1:250), Abcam, Cambridge, MA, mouse anti-mitofusin 1 (MFN1) monoclonal antibody (1:100), Abcam, Cambridge, MA, mouse anti-dynamin-related protein 1 (DRP1) monoclonal antibody (1:100), Abcam, Cambridge, MA, rabbit anti-very long-chain acyl-CoA dehydrogenase (VLCAD) antiserum (1:1,000), Cocalico Biologicals Inc., PA, rabbit anti-voltage-dependent anion channel 1 (VDAC1) monoclonal antibody (1:1,000), Abcam, Cambridge, MA, mouse anti-glucose-related protein 75 (Grp75) monoclonal antibody (1:250), Abcam, Cambridge, MA, rabbit anti-glucose-related protein 78 (Grp78) polyclonal antibody (1:250), Abcam, Cambridge, MA, mouse anti-DNA damage inducible transcript 3 (DDIT3) monoclonal antibody (1:250), Abcam, Cambridge, MA, goat anti-inositol 1,4,5-trisphosphate receptor 3 (IP3R) polyclonal antibody (1:50), Santa Cruz Biotechnology, Dallas, TX, or IgG-HRP conjugated antibody, Bio-Rad, Hercules, CA.

Techniques: Cell Culture, Incubation

Journal: Scientific Reports

Article Title: Evaluation of mitochondrial bioenergetics, dynamics, endoplasmic reticulum-mitochondria crosstalk, and reactive oxygen species in fibroblasts from patients with complex I deficiency

doi: 10.1038/s41598-018-19543-3

Figure Lengend Snippet: Superoxide production in ND6 deficient, NDUFV1 deficient and ACAD9 deficient fibroblasts cultured in media with ( A ) or without glucose for 48 h ( B ). Data are means ± SD. *P < 0.05, ***P < 0.001, compared to wild type (WT) cells ( t test for unpaired samples) ( A and B ).

Article Snippet: Following electrophoresis, the gel was blotted onto a nitrocellulose membrane, which was incubated with rabbit anti-ND6 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-NDUFV1 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-ACAD9 antiserum (1:500), Cocalico Biologicals Inc., PA, rodent anti-total OXPHOS cocktail antibody (1:250), Abcam, Cambridge, MA, mouse anti-mitofusin 1 (MFN1) monoclonal antibody (1:100), Abcam, Cambridge, MA, mouse anti-dynamin-related protein 1 (DRP1) monoclonal antibody (1:100), Abcam, Cambridge, MA, rabbit anti-very long-chain acyl-CoA dehydrogenase (VLCAD) antiserum (1:1,000), Cocalico Biologicals Inc., PA, rabbit anti-voltage-dependent anion channel 1 (VDAC1) monoclonal antibody (1:1,000), Abcam, Cambridge, MA, mouse anti-glucose-related protein 75 (Grp75) monoclonal antibody (1:250), Abcam, Cambridge, MA, rabbit anti-glucose-related protein 78 (Grp78) polyclonal antibody (1:250), Abcam, Cambridge, MA, mouse anti-DNA damage inducible transcript 3 (DDIT3) monoclonal antibody (1:250), Abcam, Cambridge, MA, goat anti-inositol 1,4,5-trisphosphate receptor 3 (IP3R) polyclonal antibody (1:50), Santa Cruz Biotechnology, Dallas, TX, or IgG-HRP conjugated antibody, Bio-Rad, Hercules, CA.

Techniques: Cell Culture

Journal: Scientific Reports

Article Title: Evaluation of mitochondrial bioenergetics, dynamics, endoplasmic reticulum-mitochondria crosstalk, and reactive oxygen species in fibroblasts from patients with complex I deficiency

doi: 10.1038/s41598-018-19543-3

Figure Lengend Snippet: Inositol 1,4,5-trisphosphate receptor 3 (IP3R), glucose-related protein 75 (Grp75), voltage-dependent anion channel 1 (VDAC1), glucose-related protein 78 (Grp78) and DNA damage inducible transcript (DDIT3) protein content in cell lysates prepared from ND6 deficient and ACAD9 deficient fibroblasts cultured in media with glucose Cell viability ( C ) and apoptosis ( D and E ) in ND6 deficient and ACAD9 deficient fibroblasts cultured in media with or without glucose for 48 hr. Data are means ± SD. *P < 0.05, **P < 0.01, ****P < 0.0001, compared to wild type (WT) cells ( t test for unpaired samples).

Article Snippet: Following electrophoresis, the gel was blotted onto a nitrocellulose membrane, which was incubated with rabbit anti-ND6 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-NDUFV1 polyclonal antibody (1:100), Santa Cruz Biotechnology, Dallas, TX, rabbit anti-ACAD9 antiserum (1:500), Cocalico Biologicals Inc., PA, rodent anti-total OXPHOS cocktail antibody (1:250), Abcam, Cambridge, MA, mouse anti-mitofusin 1 (MFN1) monoclonal antibody (1:100), Abcam, Cambridge, MA, mouse anti-dynamin-related protein 1 (DRP1) monoclonal antibody (1:100), Abcam, Cambridge, MA, rabbit anti-very long-chain acyl-CoA dehydrogenase (VLCAD) antiserum (1:1,000), Cocalico Biologicals Inc., PA, rabbit anti-voltage-dependent anion channel 1 (VDAC1) monoclonal antibody (1:1,000), Abcam, Cambridge, MA, mouse anti-glucose-related protein 75 (Grp75) monoclonal antibody (1:250), Abcam, Cambridge, MA, rabbit anti-glucose-related protein 78 (Grp78) polyclonal antibody (1:250), Abcam, Cambridge, MA, mouse anti-DNA damage inducible transcript 3 (DDIT3) monoclonal antibody (1:250), Abcam, Cambridge, MA, goat anti-inositol 1,4,5-trisphosphate receptor 3 (IP3R) polyclonal antibody (1:50), Santa Cruz Biotechnology, Dallas, TX, or IgG-HRP conjugated antibody, Bio-Rad, Hercules, CA.

Techniques: Cell Culture