- PA1-41082 - Invitrogen Antibodies FIS1 antibody

Submitted references Cells Lacking PA200 Adapt to Mitochondrial Dysfunction by Enhancing Glycolysis via Distinct Opa1 Processing.

Douida A, Batista F, Boto P, Regdon Z, Robaszkiewicz A, Tar K
International journal of molecular sciences 2021 Feb 5;22(4)

Lipids activate skeletal muscle mitochondrial fission and quality control networks to induce insulin resistance in humans.

Axelrod CL, Fealy CE, Erickson ML, Davuluri G, Fujioka H, Dantas WS, Huang E, Pergola K, Mey JT, King WT, Mulya A, Hsia D, Burguera B, Tandler B, Hoppel CL, Kirwan JP
Metabolism: clinical and experimental 2021 Aug;121:154803

Downregulation of Drp1 and Fis1 Inhibits Mitochondrial Fission and Prevents High Glucose-Induced Apoptosis in Retinal Endothelial Cells.

Kim D, Sankaramoorthy A, Roy S
Cells 2020 Jul 10;9(7)

Abnormal Mitochondria in a Non-human Primate Model of MPTP-induced Parkinson's Disease: Drp1 and CDK5/p25 Signaling.

Park J, Seo J, Won J, Yeo HG, Ahn YJ, Kim K, Jin YB, Koo BS, Lim KS, Jeong KJ, Kang P, Lee HY, Baek SH, Jeon CY, Hong JJ, Huh JW, Kim YH, Park SJ, Kim SU, Lee DS, Lee SR, Lee Y
Experimental neurobiology 2019 Jun;28(3):414-424

Loss of succinyl-CoA synthase ADP-forming β subunit disrupts mtDNA stability and mitochondrial dynamics in neurons.

Zhao Y, Tian J, Sui S, Yuan X, Chen H, Qu C, Du Y, Guo L, Du H
Scientific reports 2017 Aug 2;7(1):7169

Autophagy signaling in skeletal muscle of infarcted rats.

Jannig PR, Moreira JB, Bechara LR, Bozi LH, Bacurau AV, Monteiro AW, Dourado PM, Wisløff U, Brum PC
PloS one 2014;9(1):e85820

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown validation of FIS1 in A) human HeLa and B) mouse NIH 3T3 lysate. Samples were incubated in FIS1 polyclonal (Product # PA1-41082) using a dilution of 1:1000. B. Specificity validation of the Fis1 antibody. Fis1 was analyzed in M17 cells stably transfected with vector only, Fis1 RNAi, Fis1 (Fis1wt OE) or left untransfected (negative control). The western blot data showed that M17 cells expressed Fis1 (Vector, Untransfected control) which could be knocked down by Fis1 RNAi. Cells transfected with Fis1 overexpressed Fis1. M17 is a human neuroblastoma cell line.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Effect of Fis1/Drp1 siRNA on high glucose (HG)-induced overexpression of Fis1 and Drp1 expression in rat retinal endothelial cells (RRECs). ( A ) Representative Western blot (WB) image shows Fis1 and Drp1 expression is significantly upregulated in cells grown in HG medium. In parallel, cells grown in HG and transfected with Fis1 siRNA, Drp1 siRNA, or both siRNAs showed corresponding decreases in Fis1 or Drp1 expression. Graphical illustration of cumulative data shows Drp1 ( B ) and Fis1 ( C ) are significantly upregulated in cells grown in HG condition and reduced in cells transfected with Drp1 and Fis1 siRNAs. Data are expressed as mean +- SD. * p < 0.05. n = 6.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 Mitophagy and mitochondrial fission in soleus muscle. Soleus Bnip3 ( A ), DRP1 ( B ) and Fis1 ( C ) protein levels, and representative immunoblots ( D ) in Sham and MI groups. Correlation between soleus BNIP3 mRNA levels and distance run in a graded treadmill exercise test ( E , Sham n = 8, MI n = 10). Data presented as mean +- SEM. The number of animals in each analysis is shown within the bars.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 7 Mitophagy and mitochondrial fission in plantaris muscle. Plantaris Bnip3 ( A ), DRP1 ( B ) and Fis1 ( C ) protein levels, and representative immunoblots ( D ) in Sham and MI groups. Correlation between plantaris BNIP3 mRNA levels and distance run in a graded treadmill exercise test ( E , Sham n = 8, MI n = 10). Data presented as mean +- SEM. *indicates p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Loss of SCS A-beta suppresses the expression of mitochondrial fusion and fission proteins. The expression levels of Mfn2 ( a ), OPA1 ( b ), FIS1 ( c ), Dlp1 ( d ) and pDlp1S616 ( e ) were measured by using immunoblotting. The lower panels are representative immunoblots. The data were collected from 3 independent experiments.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Effect of Fis1/Drp1 siRNA on high glucose (HG)-induced overexpression of Fis1 and Drp1 expression in rat retinal endothelial cells (RRECs). ( A ) Representative Western blot (WB) image shows Fis1 and Drp1 expression is significantly upregulated in cells grown in HG medium. In parallel, cells grown in HG and transfected with Fis1 siRNA, Drp1 siRNA, or both siRNAs showed corresponding decreases in Fis1 or Drp1 expression. Graphical illustration of cumulative data shows Drp1 ( B ) and Fis1 ( C ) are significantly upregulated in cells grown in HG condition and reduced in cells transfected with Drp1 and Fis1 siRNAs. Data are expressed as mean +- SD. * p < 0.05. n = 6.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 Effect of reducing HG-induced overexpression of fission genes on apoptosis. ( A ) Representative WB image shows cleaved caspase-3 and Bax expression are significantly upregulated in cells grown in HG medium. In contrast, cells grown in HG medium and transfected with Fis1 siRNA, Drp1 siRNA, or both siRNAs exhibit reduced levels of these pro-apoptotic genes. Graphical illustration of cumulative data shows ( B ) cleaved caspase-3 and ( C ) Bax levels are downregulated in cells transfected with Fis1 siRNA, Drp1 siRNA, or both siRNAs compared with those of cells grown in HG medium alone. Data are expressed as mean +- SD. * p < 0.05. n = 6.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 8 PA200-deficient cells exhibit low stress-induced L-OPA1 cleavage. Control and PA200-depleted cells were treated with either vehicle (DMSO) or 3 uM oligomycin for 24 h. Cells were lysed with RIPA buffer and an equal amount of protein was separated using SDS-PAGE and analyzed by western blot. ( A ) The protein levels of mitochondrial fission and fusion proteins were examined using anti-Drp1, anti-Fis1, anti-Mfn1, anti-Mfn2, and anti-Mff antibodies. beta-actin was used as an internal loading control. ( B ) Mitochondrial fusion protein levels were examined using an anti-Opa1 antibody. Representative images of western blots and statistical analysis of the relative protein levels are shown. Images were taken using a ChemiDoc imager, and the pixel intensity was quantified and normalized to the internal control, beta-actin. The ratio of L-Opa1 to S-Opa1 was determined. Data are presented as mean values +- SD ( n = 4). Groups were compared using two-way ANOVA and t -tests. (** indicates p < 0.01, *** indicates p < 0.001, ## indicates p < 0.01) ( C ) IM peptidase OMA1 protein levels were examined using an anti-OMA1 antibody. Representative images of western blot and statistical analysis of the relative protein level are shown. Images were taken using a ChemiDoc imager, and the pixel intensity was quantified and normalized to the internal control, beta-actin. Data are presented as mean values +- SD ( n = 4). Statistical analysis was performed using two-way ANOVA. (* indicates p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 Expression level of proteins involved in mitochondrial dynamics. (A) The levels of the mitochondrial fission proteins, p-Drp1(S616), Drp1, and Fis1, and (B) the mitochondrial fusion proteins, Mfn1, Mfn2, and Opa1, in the substantia nigra of saline- or MPTP-injected cynomolgus monkeys were identified using western blot analysis. Drp1 was used as the loading control for p-Drp1(S616). The data are presented as mean values+-SD (n=2). ** denotes p

PA1-41082

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