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Santa Cruz Biotechnology subunit 8a tim
Fig. 1 Characterization of rat sciatic nerve- derived fractions. Figure shows the semiquantitative WB analysis of typical myelin proteins, as MBP, PMP22, myelin protein zero (MPZ) and typical mitochon- drial proteins, as adenosine nucleotide translocase (ANT), inner membrane tran- slocase, subunit <t>8A</t> <t>(TIM),</t> outer mem- brane translocase, subunit 20 (TOM) and Adenylate kinase, isoform 3 (AK3) in mitochondria-enriched fractions (lane 1), sciatic nerve homogenate (lane 2) and isolated myelin vesicles (IMV, lane 3) from 12-month-old wild type (Wt) rats. Panel (a) reports the protein pattern stained with Colloidal Blue Coomassie. In panels (b–h) WB analysis against MBP, PMP22, MPZ, ANT,TIM, TOM and AK3 is shown, respectively. The densitometric analysis of WB signals is reported in Panel (i). To further assess the absence of contami- nation from mitochondrial matrix we checked for AK3 activity in PNS IMV. AK3 activity was strongly impaired in SN, and undetectable in IMV compared to MF (Panel j). Panel (k) shows the amplifi- cation of a specific mitochondrial DNA sequence. The signal is present only in the mitochondrial sample, used as positive control, and absent in IMV. Each Panel is representative of at least 10 different experiments.
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Image Search Results


Fig. 1 Characterization of rat sciatic nerve- derived fractions. Figure shows the semiquantitative WB analysis of typical myelin proteins, as MBP, PMP22, myelin protein zero (MPZ) and typical mitochon- drial proteins, as adenosine nucleotide translocase (ANT), inner membrane tran- slocase, subunit 8A (TIM), outer mem- brane translocase, subunit 20 (TOM) and Adenylate kinase, isoform 3 (AK3) in mitochondria-enriched fractions (lane 1), sciatic nerve homogenate (lane 2) and isolated myelin vesicles (IMV, lane 3) from 12-month-old wild type (Wt) rats. Panel (a) reports the protein pattern stained with Colloidal Blue Coomassie. In panels (b–h) WB analysis against MBP, PMP22, MPZ, ANT,TIM, TOM and AK3 is shown, respectively. The densitometric analysis of WB signals is reported in Panel (i). To further assess the absence of contami- nation from mitochondrial matrix we checked for AK3 activity in PNS IMV. AK3 activity was strongly impaired in SN, and undetectable in IMV compared to MF (Panel j). Panel (k) shows the amplifi- cation of a specific mitochondrial DNA sequence. The signal is present only in the mitochondrial sample, used as positive control, and absent in IMV. Each Panel is representative of at least 10 different experiments.

Journal: Journal of neurochemistry

Article Title: Oxydative phosphorylation in sciatic nerve myelin and its impairment in a model of dysmyelinating peripheral neuropathy.

doi: 10.1111/jnc.12253

Figure Lengend Snippet: Fig. 1 Characterization of rat sciatic nerve- derived fractions. Figure shows the semiquantitative WB analysis of typical myelin proteins, as MBP, PMP22, myelin protein zero (MPZ) and typical mitochon- drial proteins, as adenosine nucleotide translocase (ANT), inner membrane tran- slocase, subunit 8A (TIM), outer mem- brane translocase, subunit 20 (TOM) and Adenylate kinase, isoform 3 (AK3) in mitochondria-enriched fractions (lane 1), sciatic nerve homogenate (lane 2) and isolated myelin vesicles (IMV, lane 3) from 12-month-old wild type (Wt) rats. Panel (a) reports the protein pattern stained with Colloidal Blue Coomassie. In panels (b–h) WB analysis against MBP, PMP22, MPZ, ANT,TIM, TOM and AK3 is shown, respectively. The densitometric analysis of WB signals is reported in Panel (i). To further assess the absence of contami- nation from mitochondrial matrix we checked for AK3 activity in PNS IMV. AK3 activity was strongly impaired in SN, and undetectable in IMV compared to MF (Panel j). Panel (k) shows the amplifi- cation of a specific mitochondrial DNA sequence. The signal is present only in the mitochondrial sample, used as positive control, and absent in IMV. Each Panel is representative of at least 10 different experiments.

Article Snippet: Primary antibodies (Ab) were: monoclonal antibody against myelin protein zero (MPZ) (P07 extracellular domain, Astexx Ltd. & Co. KEG, Graz, Austria); polyclonal antibodies against: adenosine nucleotide translocase (ANT) (Q-18, sc-9300 Santa Cruz Biotechnology, Santa Cruz, CA, USA); mitochondrial import inner membrane translocase, subunit 8A (TIM) (N-20, sc-17052, Santa Cruz Biotechnology); mitochondrial import outer membrane translocase, subunit 20 (TOM) (C-20, sc-11021, Santa Cruz Biotechnology); FoF1-ATP synthase a-subunit (A9585, Sigma-Aldrich, St. Louis, MO, USA); NADH-ubiquinone oxidoreductase subunit (ND4L) (H-94, sc-20665, Santa Cruz Biotechnology), and cytochrome c oxydase, subunit II (COX II) (Q-18, sc-23983, Santa Cruz Biotechnology).

Techniques: Derivative Assay, Membrane, Isolation, Staining, Activity Assay, Sequencing, Positive Control