mouse polyclonal park2 parkin antibody Search Results


park2  (Novus Biologicals)
90
Novus Biologicals park2
SIRT5 controls ammonia-induced mitophagy and mitochondrial fusion proteins. ( A ) Upper panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with 100 nM bafilomycinA 1 , BPTES, dimethyl-α-ketoglutarate, NH 4 Cl or grown without L -glutamine for 17 h and processed. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Middle panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. PINK1 and <t>PARK2</t> levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05. ( B ) Upper panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05.
Park2, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 90 stars, based on 1 article reviews
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phospho ser65 park2  (Bioss)
94
Bioss phospho ser65 park2
SIRT5 controls ammonia-induced mitophagy and mitochondrial fusion proteins. ( A ) Upper panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with 100 nM bafilomycinA 1 , BPTES, dimethyl-α-ketoglutarate, NH 4 Cl or grown without L -glutamine for 17 h and processed. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Middle panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. PINK1 and <t>PARK2</t> levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05. ( B ) Upper panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05.
Phospho Ser65 Park2, supplied by Bioss, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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park2 parkin polyclonal antibody  (Proteintech)
96
Proteintech park2 parkin polyclonal antibody
SIRT5 controls ammonia-induced mitophagy and mitochondrial fusion proteins. ( A ) Upper panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with 100 nM bafilomycinA 1 , BPTES, dimethyl-α-ketoglutarate, NH 4 Cl or grown without L -glutamine for 17 h and processed. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Middle panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. PINK1 and <t>PARK2</t> levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05. ( B ) Upper panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05.
Park2 Parkin Polyclonal Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/park2 parkin polyclonal antibody/product/Proteintech
Average 96 stars, based on 1 article reviews
park2 parkin polyclonal antibody - by Bioz Stars, 2026-03
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mouse anti prkn parkin  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc mouse anti prkn parkin
SIRT5 controls ammonia-induced mitophagy and mitochondrial fusion proteins. ( A ) Upper panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with 100 nM bafilomycinA 1 , BPTES, dimethyl-α-ketoglutarate, NH 4 Cl or grown without L -glutamine for 17 h and processed. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Middle panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. PINK1 and <t>PARK2</t> levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05. ( B ) Upper panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05.
Mouse Anti Prkn Parkin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse anti prkn parkin - by Bioz Stars, 2026-03
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anti prkn  (Santa Cruz Biotechnology)
96
Santa Cruz Biotechnology anti prkn
SIRT5 controls ammonia-induced mitophagy and mitochondrial fusion proteins. ( A ) Upper panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with 100 nM bafilomycinA 1 , BPTES, dimethyl-α-ketoglutarate, NH 4 Cl or grown without L -glutamine for 17 h and processed. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Middle panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. PINK1 and <t>PARK2</t> levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05. ( B ) Upper panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05.
Anti Prkn, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti prkn/product/Santa Cruz Biotechnology
Average 96 stars, based on 1 article reviews
anti prkn - by Bioz Stars, 2026-03
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prkn park2  (Cell Signaling Technology Inc)
97
Cell Signaling Technology Inc prkn park2
BNIP3L relates to mitophagy deficiency in cerebral ischemia. (A) Mice were infected with AAV-GFP-Bnip3l or AAV-GFThe mice were subjected to pMCAO for 4 h, then the protein levels of GFP, SQSTM1, LC3B, COX4I1 and TOMM20 in brain tissues were determined by western blot. Empty arrows indicate exogenous BNIP3L. Duplicate lanes are shown for each grou(B) Semi-quantitative analyses of SQSTM1, LC3B, COX4I1 and TOMM20 protein levels are shown (n = 3 mice for each group). (C) Primary neurons were subjected to OGD for 1 h. The expression of BNIP3L and TOMM20 were detected by western blot. Semi-quantitative analyses are shown (n = 3 from independent experiments). (D) The neurons overexpression Flag-BNIP3L were subjected to OGD for 1 h, and the expression of Flag-BNIP3L and TOMM20 were detected by western blot (n = 3 from independent experiments). (E) Mice were subjected to 1, 3 or 6 h of pMCAO. The BNIP3L, <t>PRKN,</t> FUNDC1, BCL2L13, PHB2, SQSTM1, LC3B, COX4I1 and TOMM20 protein levels were determined by western blot. Black arrows indicate endogenous BNIP3L. Duplicate lanes are shown for each grou(F-I) Analyses of the correlations between BNIP3L, PRKN, FUNDC1, BLC2L13, PHB2, and mitochondrial proteins are shown (n = 3 mice for each group). Data are expressed as mean ± SEM. Statistical comparisons were performed as follows: one-way ANOVA for (B and F-I) and t-test for (C and D). *P < 0.05; **P < 0.01; ***P < 0.001; n.s. vs. the indicated group
Prkn Park2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti parkin  (Novus Biologicals)
92
Novus Biologicals rabbit anti parkin
BNIP3L relates to mitophagy deficiency in cerebral ischemia. (A) Mice were infected with AAV-GFP-Bnip3l or AAV-GFThe mice were subjected to pMCAO for 4 h, then the protein levels of GFP, SQSTM1, LC3B, COX4I1 and TOMM20 in brain tissues were determined by western blot. Empty arrows indicate exogenous BNIP3L. Duplicate lanes are shown for each grou(B) Semi-quantitative analyses of SQSTM1, LC3B, COX4I1 and TOMM20 protein levels are shown (n = 3 mice for each group). (C) Primary neurons were subjected to OGD for 1 h. The expression of BNIP3L and TOMM20 were detected by western blot. Semi-quantitative analyses are shown (n = 3 from independent experiments). (D) The neurons overexpression Flag-BNIP3L were subjected to OGD for 1 h, and the expression of Flag-BNIP3L and TOMM20 were detected by western blot (n = 3 from independent experiments). (E) Mice were subjected to 1, 3 or 6 h of pMCAO. The BNIP3L, <t>PRKN,</t> FUNDC1, BCL2L13, PHB2, SQSTM1, LC3B, COX4I1 and TOMM20 protein levels were determined by western blot. Black arrows indicate endogenous BNIP3L. Duplicate lanes are shown for each grou(F-I) Analyses of the correlations between BNIP3L, PRKN, FUNDC1, BLC2L13, PHB2, and mitochondrial proteins are shown (n = 3 mice for each group). Data are expressed as mean ± SEM. Statistical comparisons were performed as follows: one-way ANOVA for (B and F-I) and t-test for (C and D). *P < 0.05; **P < 0.01; ***P < 0.001; n.s. vs. the indicated group
Rabbit Anti Parkin, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti parkin  (Boster Bio)
93
Boster Bio rabbit anti parkin
BNIP3L relates to mitophagy deficiency in cerebral ischemia. (A) Mice were infected with AAV-GFP-Bnip3l or AAV-GFThe mice were subjected to pMCAO for 4 h, then the protein levels of GFP, SQSTM1, LC3B, COX4I1 and TOMM20 in brain tissues were determined by western blot. Empty arrows indicate exogenous BNIP3L. Duplicate lanes are shown for each grou(B) Semi-quantitative analyses of SQSTM1, LC3B, COX4I1 and TOMM20 protein levels are shown (n = 3 mice for each group). (C) Primary neurons were subjected to OGD for 1 h. The expression of BNIP3L and TOMM20 were detected by western blot. Semi-quantitative analyses are shown (n = 3 from independent experiments). (D) The neurons overexpression Flag-BNIP3L were subjected to OGD for 1 h, and the expression of Flag-BNIP3L and TOMM20 were detected by western blot (n = 3 from independent experiments). (E) Mice were subjected to 1, 3 or 6 h of pMCAO. The BNIP3L, <t>PRKN,</t> FUNDC1, BCL2L13, PHB2, SQSTM1, LC3B, COX4I1 and TOMM20 protein levels were determined by western blot. Black arrows indicate endogenous BNIP3L. Duplicate lanes are shown for each grou(F-I) Analyses of the correlations between BNIP3L, PRKN, FUNDC1, BLC2L13, PHB2, and mitochondrial proteins are shown (n = 3 mice for each group). Data are expressed as mean ± SEM. Statistical comparisons were performed as follows: one-way ANOVA for (B and F-I) and t-test for (C and D). *P < 0.05; **P < 0.01; ***P < 0.001; n.s. vs. the indicated group
Rabbit Anti Parkin, supplied by Boster Bio, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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prkn mouse monoclonal antibody  (Santa Cruz Biotechnology)
90
Santa Cruz Biotechnology prkn mouse monoclonal antibody
BNIP3L relates to mitophagy deficiency in cerebral ischemia. (A) Mice were infected with AAV-GFP-Bnip3l or AAV-GFThe mice were subjected to pMCAO for 4 h, then the protein levels of GFP, SQSTM1, LC3B, COX4I1 and TOMM20 in brain tissues were determined by western blot. Empty arrows indicate exogenous BNIP3L. Duplicate lanes are shown for each grou(B) Semi-quantitative analyses of SQSTM1, LC3B, COX4I1 and TOMM20 protein levels are shown (n = 3 mice for each group). (C) Primary neurons were subjected to OGD for 1 h. The expression of BNIP3L and TOMM20 were detected by western blot. Semi-quantitative analyses are shown (n = 3 from independent experiments). (D) The neurons overexpression Flag-BNIP3L were subjected to OGD for 1 h, and the expression of Flag-BNIP3L and TOMM20 were detected by western blot (n = 3 from independent experiments). (E) Mice were subjected to 1, 3 or 6 h of pMCAO. The BNIP3L, <t>PRKN,</t> FUNDC1, BCL2L13, PHB2, SQSTM1, LC3B, COX4I1 and TOMM20 protein levels were determined by western blot. Black arrows indicate endogenous BNIP3L. Duplicate lanes are shown for each grou(F-I) Analyses of the correlations between BNIP3L, PRKN, FUNDC1, BLC2L13, PHB2, and mitochondrial proteins are shown (n = 3 mice for each group). Data are expressed as mean ± SEM. Statistical comparisons were performed as follows: one-way ANOVA for (B and F-I) and t-test for (C and D). *P < 0.05; **P < 0.01; ***P < 0.001; n.s. vs. the indicated group
Prkn Mouse Monoclonal Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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prkn mouse monoclonal antibody - by Bioz Stars, 2026-03
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park2 parkin monoclonal antibody  (Proteintech)
96
Proteintech park2 parkin monoclonal antibody
BNIP3L relates to mitophagy deficiency in cerebral ischemia. (A) Mice were infected with AAV-GFP-Bnip3l or AAV-GFThe mice were subjected to pMCAO for 4 h, then the protein levels of GFP, SQSTM1, LC3B, COX4I1 and TOMM20 in brain tissues were determined by western blot. Empty arrows indicate exogenous BNIP3L. Duplicate lanes are shown for each grou(B) Semi-quantitative analyses of SQSTM1, LC3B, COX4I1 and TOMM20 protein levels are shown (n = 3 mice for each group). (C) Primary neurons were subjected to OGD for 1 h. The expression of BNIP3L and TOMM20 were detected by western blot. Semi-quantitative analyses are shown (n = 3 from independent experiments). (D) The neurons overexpression Flag-BNIP3L were subjected to OGD for 1 h, and the expression of Flag-BNIP3L and TOMM20 were detected by western blot (n = 3 from independent experiments). (E) Mice were subjected to 1, 3 or 6 h of pMCAO. The BNIP3L, <t>PRKN,</t> FUNDC1, BCL2L13, PHB2, SQSTM1, LC3B, COX4I1 and TOMM20 protein levels were determined by western blot. Black arrows indicate endogenous BNIP3L. Duplicate lanes are shown for each grou(F-I) Analyses of the correlations between BNIP3L, PRKN, FUNDC1, BLC2L13, PHB2, and mitochondrial proteins are shown (n = 3 mice for each group). Data are expressed as mean ± SEM. Statistical comparisons were performed as follows: one-way ANOVA for (B and F-I) and t-test for (C and D). *P < 0.05; **P < 0.01; ***P < 0.001; n.s. vs. the indicated group
Park2 Parkin Monoclonal Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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park2 parkin monoclonal antibody - by Bioz Stars, 2026-03
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mouse anti-prkn  (Millipore)
90
Millipore mouse anti-prkn
Complete loss of PINK1 in mouse brain stabilizes <t>PRKN</t> protein while partial loss dampens p-S65-Ub levels. PRKN and p-S65-Ub levels were determined at basal conditions in hemibrain lysates from mice with the following genotypes: WT ( n = 29), heterozygous Prkn +/- ( n = 23), homozygous prkn -/- ( n = 29), heterozygous Pink1 +/- ( n = 15), homozygous pink1 -/- ( n = 22). For better comparison, mice were grouped and analyzed by genotype: WT, prkn -/- , pink1 -/- (left); WT, Prkn +/- , prkn -/- (middle); and WT, Pink1 +/- , pink1 -/- (right). (A) Representative western blots obtained with three different anti-PRKN antibodies <t>(Prk8,</t> 2132, and 5C3) are shown together with the loading control GAPDH. An open arrowhead labels the truncated PRKN-EGFP fusion protein produced in the prkn -/- samples. (B) PRKN (Prk8) protein levels were assessed by densitometry with data points shown as ratio PRKN divided by GAPDH (median ± interquartile range [IQR]). Quantification of PRKN protein using the other two antibodies can be found in Fig. S1A and B. (C) p-S65-Ub levels were quantified by sandwich ELISA with data points shown as median ± IQR. Data was analyzed by using a Kruskal-Wallis test combined with Dunn’s multiple comparison test (***, p < 0.0005; *, p < 0.05). Comparisons to the WT are shown on top of data points, while comparisons between other genotypes are indicated by brackets.
Mouse Anti Prkn, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti th  (Novus Biologicals)
94
Novus Biologicals rabbit anti th
Complete loss of PINK1 in mouse brain stabilizes <t>PRKN</t> protein while partial loss dampens p-S65-Ub levels. PRKN and p-S65-Ub levels were determined at basal conditions in hemibrain lysates from mice with the following genotypes: WT ( n = 29), heterozygous Prkn +/- ( n = 23), homozygous prkn -/- ( n = 29), heterozygous Pink1 +/- ( n = 15), homozygous pink1 -/- ( n = 22). For better comparison, mice were grouped and analyzed by genotype: WT, prkn -/- , pink1 -/- (left); WT, Prkn +/- , prkn -/- (middle); and WT, Pink1 +/- , pink1 -/- (right). (A) Representative western blots obtained with three different anti-PRKN antibodies <t>(Prk8,</t> 2132, and 5C3) are shown together with the loading control GAPDH. An open arrowhead labels the truncated PRKN-EGFP fusion protein produced in the prkn -/- samples. (B) PRKN (Prk8) protein levels were assessed by densitometry with data points shown as ratio PRKN divided by GAPDH (median ± interquartile range [IQR]). Quantification of PRKN protein using the other two antibodies can be found in Fig. S1A and B. (C) p-S65-Ub levels were quantified by sandwich ELISA with data points shown as median ± IQR. Data was analyzed by using a Kruskal-Wallis test combined with Dunn’s multiple comparison test (***, p < 0.0005; *, p < 0.05). Comparisons to the WT are shown on top of data points, while comparisons between other genotypes are indicated by brackets.
Rabbit Anti Th, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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SIRT5 controls ammonia-induced mitophagy and mitochondrial fusion proteins. ( A ) Upper panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with 100 nM bafilomycinA 1 , BPTES, dimethyl-α-ketoglutarate, NH 4 Cl or grown without L -glutamine for 17 h and processed. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Middle panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. PINK1 and PARK2 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05. ( B ) Upper panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05.

Journal: Autophagy

Article Title: SIRT5 regulation of ammonia-induced autophagy and mitophagy

doi: 10.1080/15548627.2015.1009778

Figure Lengend Snippet: SIRT5 controls ammonia-induced mitophagy and mitochondrial fusion proteins. ( A ) Upper panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with 100 nM bafilomycinA 1 , BPTES, dimethyl-α-ketoglutarate, NH 4 Cl or grown without L -glutamine for 17 h and processed. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Middle panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. PINK1 and PARK2 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: MDA-MB-231 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05. ( B ) Upper panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain mitochondrial fractions. Alternatively, WT, SIRT5 + and SIRT5 − cells were treated with NH 4 Cl for 17 h. BNIP3 expression levels were determined by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. COX4I1 was used as a loading control. *Significantly different from WT cells. Significance was set at P < 0.05. Lower panel: C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5 + and SIRT5 − clones were processed to obtain whole cellular extracts. MFN2 and OPA1 levels were assessed by western blot as indicated under Materials and Methods. Densitometric analysis of the gels was performed as described under Materials and Methods. ACTB was used as loading control. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05.

Article Snippet: Antibodies used: SIRT5 (Abcam, 13697), MAP1LC3B (Novus Biologicals, NB600-1384), GABARAP and GABARAPL2 (MBL International Corporation, PM037 and PM038, respectively), SQSTM1 (Santa Cruz Biotechnology, sc-48402), BNIP3 (Santa Cruz Biotechnology, sc-56167), MFN2 (Abnova, H00009927-M07), OPA1 (Novus Biologicals, H00004976-D01P), PINK1 (Santa Cruz Biotechnology, sc-33796), PARK2 (Santa Cruz Biotechnology, sc-32282), GLS (Novus Biologicals, H00002744-M01), GLUL (Novus Biologicals, H00002752-M02), GLUD1 (Santa Cruz Biotechnology, sc-160382), SIRT3 (Cell Signaling Technology, 2627), succinyl lysine (PTM-Biolabs, PTM 419), acetylated lysine (Cell Signaling Technology, 9447), ACTB (Santa Cruz Biotechnology, sc-8432), COX4I1 (Santa Cruz Biotechnology, sc-69359).

Techniques: Clone Assay, Expressing, Western Blot, Control

BNIP3L relates to mitophagy deficiency in cerebral ischemia. (A) Mice were infected with AAV-GFP-Bnip3l or AAV-GFThe mice were subjected to pMCAO for 4 h, then the protein levels of GFP, SQSTM1, LC3B, COX4I1 and TOMM20 in brain tissues were determined by western blot. Empty arrows indicate exogenous BNIP3L. Duplicate lanes are shown for each grou(B) Semi-quantitative analyses of SQSTM1, LC3B, COX4I1 and TOMM20 protein levels are shown (n = 3 mice for each group). (C) Primary neurons were subjected to OGD for 1 h. The expression of BNIP3L and TOMM20 were detected by western blot. Semi-quantitative analyses are shown (n = 3 from independent experiments). (D) The neurons overexpression Flag-BNIP3L were subjected to OGD for 1 h, and the expression of Flag-BNIP3L and TOMM20 were detected by western blot (n = 3 from independent experiments). (E) Mice were subjected to 1, 3 or 6 h of pMCAO. The BNIP3L, PRKN, FUNDC1, BCL2L13, PHB2, SQSTM1, LC3B, COX4I1 and TOMM20 protein levels were determined by western blot. Black arrows indicate endogenous BNIP3L. Duplicate lanes are shown for each grou(F-I) Analyses of the correlations between BNIP3L, PRKN, FUNDC1, BLC2L13, PHB2, and mitochondrial proteins are shown (n = 3 mice for each group). Data are expressed as mean ± SEM. Statistical comparisons were performed as follows: one-way ANOVA for (B and F-I) and t-test for (C and D). *P < 0.05; **P < 0.01; ***P < 0.001; n.s. vs. the indicated group

Journal: Autophagy

Article Title: BNIP3L/NIX degradation leads to mitophagy deficiency in ischemic brains

doi: 10.1080/15548627.2020.1802089

Figure Lengend Snippet: BNIP3L relates to mitophagy deficiency in cerebral ischemia. (A) Mice were infected with AAV-GFP-Bnip3l or AAV-GFThe mice were subjected to pMCAO for 4 h, then the protein levels of GFP, SQSTM1, LC3B, COX4I1 and TOMM20 in brain tissues were determined by western blot. Empty arrows indicate exogenous BNIP3L. Duplicate lanes are shown for each grou(B) Semi-quantitative analyses of SQSTM1, LC3B, COX4I1 and TOMM20 protein levels are shown (n = 3 mice for each group). (C) Primary neurons were subjected to OGD for 1 h. The expression of BNIP3L and TOMM20 were detected by western blot. Semi-quantitative analyses are shown (n = 3 from independent experiments). (D) The neurons overexpression Flag-BNIP3L were subjected to OGD for 1 h, and the expression of Flag-BNIP3L and TOMM20 were detected by western blot (n = 3 from independent experiments). (E) Mice were subjected to 1, 3 or 6 h of pMCAO. The BNIP3L, PRKN, FUNDC1, BCL2L13, PHB2, SQSTM1, LC3B, COX4I1 and TOMM20 protein levels were determined by western blot. Black arrows indicate endogenous BNIP3L. Duplicate lanes are shown for each grou(F-I) Analyses of the correlations between BNIP3L, PRKN, FUNDC1, BLC2L13, PHB2, and mitochondrial proteins are shown (n = 3 mice for each group). Data are expressed as mean ± SEM. Statistical comparisons were performed as follows: one-way ANOVA for (B and F-I) and t-test for (C and D). *P < 0.05; **P < 0.01; ***P < 0.001; n.s. vs. the indicated group

Article Snippet: The following primary antibodies were used: ATG7 (1:1,000; Cell Signaling Technology, 8558 T), BCL2L13 (1:1000; Proteintech; 16,612-1-AP), BNIP3L (1:1,000; Cell Signaling Technology, 12396s), COX4I1 (1:1,000; Cell Signaling Technology, 4850), Flag (1:1,000; Cell Signaling Technology, 14793s), FUNDC1 (1:1,000; Abcam, ab74834), GAPDH (1:3,000; KangChen, KC-5G4), GFP (1:1,000; MBL; 598), LC3B (1:1,000; Sigma-Aldrich, L7543), MYC (1:1,000; MBL; M192-3), PHB2/prohibitin 2 (1:1000; Proteintech; 12,295-1-AP); PRKN/PARK2 (1:1,000; Cell Signaling Technology, 2132s), SQSTM1 (1:1,000; Cell Signaling Technology, 5114), TOMM20 (1:1,000; Ambo Biotechnology, c16678), TP53 (1:1000; Cell Signaling Technology, 2524 T) and ubiquitin (1:1000, Cell Signaling Technology, 3936s).

Techniques: Infection, Western Blot, Expressing, Over Expression

Complete loss of PINK1 in mouse brain stabilizes PRKN protein while partial loss dampens p-S65-Ub levels. PRKN and p-S65-Ub levels were determined at basal conditions in hemibrain lysates from mice with the following genotypes: WT ( n = 29), heterozygous Prkn +/- ( n = 23), homozygous prkn -/- ( n = 29), heterozygous Pink1 +/- ( n = 15), homozygous pink1 -/- ( n = 22). For better comparison, mice were grouped and analyzed by genotype: WT, prkn -/- , pink1 -/- (left); WT, Prkn +/- , prkn -/- (middle); and WT, Pink1 +/- , pink1 -/- (right). (A) Representative western blots obtained with three different anti-PRKN antibodies (Prk8, 2132, and 5C3) are shown together with the loading control GAPDH. An open arrowhead labels the truncated PRKN-EGFP fusion protein produced in the prkn -/- samples. (B) PRKN (Prk8) protein levels were assessed by densitometry with data points shown as ratio PRKN divided by GAPDH (median ± interquartile range [IQR]). Quantification of PRKN protein using the other two antibodies can be found in Fig. S1A and B. (C) p-S65-Ub levels were quantified by sandwich ELISA with data points shown as median ± IQR. Data was analyzed by using a Kruskal-Wallis test combined with Dunn’s multiple comparison test (***, p < 0.0005; *, p < 0.05). Comparisons to the WT are shown on top of data points, while comparisons between other genotypes are indicated by brackets.

Journal: Autophagy

Article Title: Basal activity of PINK1 and PRKN in cell models and rodent brain

doi: 10.1080/15548627.2023.2286414

Figure Lengend Snippet: Complete loss of PINK1 in mouse brain stabilizes PRKN protein while partial loss dampens p-S65-Ub levels. PRKN and p-S65-Ub levels were determined at basal conditions in hemibrain lysates from mice with the following genotypes: WT ( n = 29), heterozygous Prkn +/- ( n = 23), homozygous prkn -/- ( n = 29), heterozygous Pink1 +/- ( n = 15), homozygous pink1 -/- ( n = 22). For better comparison, mice were grouped and analyzed by genotype: WT, prkn -/- , pink1 -/- (left); WT, Prkn +/- , prkn -/- (middle); and WT, Pink1 +/- , pink1 -/- (right). (A) Representative western blots obtained with three different anti-PRKN antibodies (Prk8, 2132, and 5C3) are shown together with the loading control GAPDH. An open arrowhead labels the truncated PRKN-EGFP fusion protein produced in the prkn -/- samples. (B) PRKN (Prk8) protein levels were assessed by densitometry with data points shown as ratio PRKN divided by GAPDH (median ± interquartile range [IQR]). Quantification of PRKN protein using the other two antibodies can be found in Fig. S1A and B. (C) p-S65-Ub levels were quantified by sandwich ELISA with data points shown as median ± IQR. Data was analyzed by using a Kruskal-Wallis test combined with Dunn’s multiple comparison test (***, p < 0.0005; *, p < 0.05). Comparisons to the WT are shown on top of data points, while comparisons between other genotypes are indicated by brackets.

Article Snippet: Membranes were blocked with 5% skim milk in TBS with 0.1% Tween (TBST) and incubated overnight with the following primary antibodies: rabbit anti-PINK1 (Cell Signaling Technology, 6946; 1:2000), mouse anti-PINK1 (Biolegend, 846201; 1:1000), mouse anti-PRKN (Millipore, MAB5521; Prk8; 1:1000–5000 for cell lysates), mouse anti-PRKN (Cell Signaling Technology, 4211; Prk8; 1:25,000 for rodent lysates]), mouse anti-PRKN (Biolegend, 865602; 5C3; 1:2,000 for rodent lysates), rabbit anti-PRKN (Cell Signaling Technology, 2132; 1:2,000 for rodent lysates), rabbit anti-p-S65-Ub (Cell Signaling Technology, 62802; 1:3000–20,000), rabbit anti-GFP (Takara/Clontech, 632460; 1:1,000 for rodent lysates) mouse anti-MFN2 (Abcam, ab56889; 1:2000), rabbit anti-TUBB3/betaIII-tubulin (Cell Signaling Technology, 5568; 1:10,000), rabbit anti-TH/tyrosine hydroxylase (Millipore, ab152, 1:1000), mouse anti-GAPDH (Meridian Life science, H86504M: 1:400,000), mouse anti-VCL/vinculin (Sigma-Aldrich, V9131; 1:500,000–1,500,000).

Techniques: Comparison, Western Blot, Produced, Sandwich ELISA

PINK1 gene dosage affects p-S65-Ub and PRKN levels in PD patients’ cells. Primary skin fibroblasts from related individuals without (QQ; n = 2), with one (QX; n = 3) or with two (XX; n = 2) mutant PINK1 Q456X alleles were either left untreated (0 h, left) or were treated for 2 h (middle) or 8 h (right) with 1 µM valinomycin (Val). Data is arranged by treatment groups (i.e., time points). (A) Cell lysates were analyzed by western blot using antibodies against PINK1, PRKN, and the loading control VCL (vinculin). (B) Protein levels of PINK1 were quantified by sandwich ELISA. (C) PRKN protein levels were derived by densitometry of the western blots and data points shown as a ratio of PRKN divided by VCL and normalized to PINK1 XX . (D) p-S65-Ub levels were also quantified by sandwich ELISA. Data is shown as the mean ± standard error of the mean of biological replicates, grouped by allele count of the Q456X mutation, and analyzed by one-way ANOVA with Tukey’s multiple comparison test (***, p < 0.0005; **, p < 0.005; *, p < 0.05). For untreated the mean of 4 technical repeats per biological sample is shown. Asterisks on top of data points indicate individual comparison to WT controls without PINK1 mutation.

Journal: Autophagy

Article Title: Basal activity of PINK1 and PRKN in cell models and rodent brain

doi: 10.1080/15548627.2023.2286414

Figure Lengend Snippet: PINK1 gene dosage affects p-S65-Ub and PRKN levels in PD patients’ cells. Primary skin fibroblasts from related individuals without (QQ; n = 2), with one (QX; n = 3) or with two (XX; n = 2) mutant PINK1 Q456X alleles were either left untreated (0 h, left) or were treated for 2 h (middle) or 8 h (right) with 1 µM valinomycin (Val). Data is arranged by treatment groups (i.e., time points). (A) Cell lysates were analyzed by western blot using antibodies against PINK1, PRKN, and the loading control VCL (vinculin). (B) Protein levels of PINK1 were quantified by sandwich ELISA. (C) PRKN protein levels were derived by densitometry of the western blots and data points shown as a ratio of PRKN divided by VCL and normalized to PINK1 XX . (D) p-S65-Ub levels were also quantified by sandwich ELISA. Data is shown as the mean ± standard error of the mean of biological replicates, grouped by allele count of the Q456X mutation, and analyzed by one-way ANOVA with Tukey’s multiple comparison test (***, p < 0.0005; **, p < 0.005; *, p < 0.05). For untreated the mean of 4 technical repeats per biological sample is shown. Asterisks on top of data points indicate individual comparison to WT controls without PINK1 mutation.

Article Snippet: Membranes were blocked with 5% skim milk in TBS with 0.1% Tween (TBST) and incubated overnight with the following primary antibodies: rabbit anti-PINK1 (Cell Signaling Technology, 6946; 1:2000), mouse anti-PINK1 (Biolegend, 846201; 1:1000), mouse anti-PRKN (Millipore, MAB5521; Prk8; 1:1000–5000 for cell lysates), mouse anti-PRKN (Cell Signaling Technology, 4211; Prk8; 1:25,000 for rodent lysates]), mouse anti-PRKN (Biolegend, 865602; 5C3; 1:2,000 for rodent lysates), rabbit anti-PRKN (Cell Signaling Technology, 2132; 1:2,000 for rodent lysates), rabbit anti-p-S65-Ub (Cell Signaling Technology, 62802; 1:3000–20,000), rabbit anti-GFP (Takara/Clontech, 632460; 1:1,000 for rodent lysates) mouse anti-MFN2 (Abcam, ab56889; 1:2000), rabbit anti-TUBB3/betaIII-tubulin (Cell Signaling Technology, 5568; 1:10,000), rabbit anti-TH/tyrosine hydroxylase (Millipore, ab152, 1:1000), mouse anti-GAPDH (Meridian Life science, H86504M: 1:400,000), mouse anti-VCL/vinculin (Sigma-Aldrich, V9131; 1:500,000–1,500,000).

Techniques: Mutagenesis, Western Blot, Sandwich ELISA, Derivative Assay, Comparison

Cell type-specific expression of PINK1 and PRKN in fibroblasts, iPSCs, and midbrain DA neurons drive p-S65-Ub levels. Primary skin fibroblasts from either a control or patient with PINK1 I368N mutation, and undifferentiated iPS cells that were generated from the same PINK1 I368N patient cells and their gene-corrected counterparts (isogenic WT), as well as DA neurons generated from the same iPSC set were treated with 1 µM valinomycin (Val) for the indicated times and harvested. (A) Representative western blots show levels of PINK1, PRKN, p-S65-Ub and MFN2 for all three cell lines. VCL (vinculin) and DA neuronal markers, TUBB3/βIII-tubulin and TH (tyrosine hydroxylase), were used as loading controls. (B) Quantification of PINK1 protein levels by sandwich ELISA. (C) Densitometric analysis of PRKN western blots shown under (A) and data points displayed as PRKN divided by VCL or PRKN divided by DA neuronal markers. (D) Quantification of p-S65-Ub levels measured by sandwich ELISA. (E) Densitometric analysis of MFN2. Relative modification of MFN2 was calculated as the ratio of upper (ubiquitinated) to lower (unmodified) MFN2 band. The mean of three independent experiments for each cell type ± SD is shown. Statistical analysis was performed by one-way ANOVA followed by Bonferroni correction. (***, p < 0.0005; **, p < 0.005; *, p < 0.05). Asterisks on top of data points indicate individual comparison to respective WT controls without PINK1 mutation.

Journal: Autophagy

Article Title: Basal activity of PINK1 and PRKN in cell models and rodent brain

doi: 10.1080/15548627.2023.2286414

Figure Lengend Snippet: Cell type-specific expression of PINK1 and PRKN in fibroblasts, iPSCs, and midbrain DA neurons drive p-S65-Ub levels. Primary skin fibroblasts from either a control or patient with PINK1 I368N mutation, and undifferentiated iPS cells that were generated from the same PINK1 I368N patient cells and their gene-corrected counterparts (isogenic WT), as well as DA neurons generated from the same iPSC set were treated with 1 µM valinomycin (Val) for the indicated times and harvested. (A) Representative western blots show levels of PINK1, PRKN, p-S65-Ub and MFN2 for all three cell lines. VCL (vinculin) and DA neuronal markers, TUBB3/βIII-tubulin and TH (tyrosine hydroxylase), were used as loading controls. (B) Quantification of PINK1 protein levels by sandwich ELISA. (C) Densitometric analysis of PRKN western blots shown under (A) and data points displayed as PRKN divided by VCL or PRKN divided by DA neuronal markers. (D) Quantification of p-S65-Ub levels measured by sandwich ELISA. (E) Densitometric analysis of MFN2. Relative modification of MFN2 was calculated as the ratio of upper (ubiquitinated) to lower (unmodified) MFN2 band. The mean of three independent experiments for each cell type ± SD is shown. Statistical analysis was performed by one-way ANOVA followed by Bonferroni correction. (***, p < 0.0005; **, p < 0.005; *, p < 0.05). Asterisks on top of data points indicate individual comparison to respective WT controls without PINK1 mutation.

Article Snippet: Membranes were blocked with 5% skim milk in TBS with 0.1% Tween (TBST) and incubated overnight with the following primary antibodies: rabbit anti-PINK1 (Cell Signaling Technology, 6946; 1:2000), mouse anti-PINK1 (Biolegend, 846201; 1:1000), mouse anti-PRKN (Millipore, MAB5521; Prk8; 1:1000–5000 for cell lysates), mouse anti-PRKN (Cell Signaling Technology, 4211; Prk8; 1:25,000 for rodent lysates]), mouse anti-PRKN (Biolegend, 865602; 5C3; 1:2,000 for rodent lysates), rabbit anti-PRKN (Cell Signaling Technology, 2132; 1:2,000 for rodent lysates), rabbit anti-p-S65-Ub (Cell Signaling Technology, 62802; 1:3000–20,000), rabbit anti-GFP (Takara/Clontech, 632460; 1:1,000 for rodent lysates) mouse anti-MFN2 (Abcam, ab56889; 1:2000), rabbit anti-TUBB3/betaIII-tubulin (Cell Signaling Technology, 5568; 1:10,000), rabbit anti-TH/tyrosine hydroxylase (Millipore, ab152, 1:1000), mouse anti-GAPDH (Meridian Life science, H86504M: 1:400,000), mouse anti-VCL/vinculin (Sigma-Aldrich, V9131; 1:500,000–1,500,000).

Techniques: Expressing, Mutagenesis, Generated, Western Blot, Sandwich ELISA, Modification, Comparison