Mouse anti fis1

Manufactured by Santa Cruz Biotechnology

Mouse anti-FIS1 is a primary antibody that specifically recognizes the FIS1 (Fission 1 Homolog) protein. FIS1 is a key regulator of mitochondrial fission and plays a crucial role in the dynamic processes of mitochondrial division and distribution within the cell.

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Anti‐Fis1 (10 citations)

2 protocols using mouse anti fis1

Western Blot Analysis of Mitochondrial Dynamics

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Human and mouse LV tissue samples were lysed in T-PER buffer (Thermo Fisher) supplemented with Halt Protease Inhibitor Cocktail (Thermo Scientific) using Qiagen Tissue Lyser II (Qiagen). Protein concentrations were determined using Pierce BCA protein assay kit (Thermo Fisher). Equal amounts of protein were loaded on 10% denaturing sodium dodecyl sulfate-polyacrylamide gels and subjected to electrophoresis, with subsequent transfer to a nitrocellulose membrane (GE Healthcare). Membranes were blocked with 1% BSA and incubated with primary antibodies, and then with a species-appropriate secondary anti-rabbit or anti-mouse fluorescently labelled IRDye secondary antibodies (Li-cor Biotechnology). Intensity of fluorescent signal was measured with Odyssey CLx imaging system (Li-cor Biotechnology) and analyzed using Fiji software (NIH). The following primary antibodies were used: rabbit anti-phospho-DRP1, rabbit anti-DRP1, rabbit anti-mitofusin-1, rabbit anti-mitofusin-2, rabbit anti-OPA1, rabbit anti-VDAC, rabbit anti-calpastatin, rabbit anti-calpain 1, rabbit anti-calpain 2, rabbit anti-OPA1 (Cell Signaling Technology), mouse anti-LC3α/β, mouse anti-parkin, mouse anti-PINK1, mouse anti-FIS1 (Santa Cruz Biotechnology), rabbit anti-NOX4 antibody (Abcam), and mouse anti-β-tubulin (Sigma-Aldrich).

Lozhkin A., Vendrov A.E., Ramos-Mondragón R., Canugovi C., Stevenson M.D., Herron T.J., Hummel S.L., Figueroa C.A., Bowles D.E., Isom L.L., Runge M.S, & Madamanchi N.R. (2022). Mitochondrial oxidative stress contributes to diastolic dysfunction through impaired mitochondrial dynamics. Redox Biology, 57, 102474.

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Western Blot Analysis of Mitochondrial Dynamics

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Protein was extracted from cortex of each rat and placed in lysis buffer to measure global Mfn1, Mfn2, Drp1, Fis1, and β-actin expression. Protein samples (30 ug each) were separated on a 10% SDS-polyacrylamide gel and transferred to polyvinylidene difluoride membranes. After blocking the nonspecific binding sites with 5% nonfat milk for 60 min, membrane was incubated overnight at 4°C with antibodies including rabbit anti-Mfn1 (Cell Signaling Technology, 1 : 1000 dilution), rabbit anti-Mfn2 (Cell Signaling Technology, 1 : 1000 dilution), rabbit anti-Drp1 (Cell Signaling Technology, 1 : 1000 dilution), mouse anti-Fis1 (Santa Cruz, 1 : 500 dilution), and mouse anti-β-actin (Cell Signaling Technology, 1 : 5000 dilution). The membranes incubated with horseradish peroxidase-conjugated secondary antibody for 45 min at room temperature and developed using an enhanced chemiluminescence detection method (Cell Signaling Technology). The relative protein expression was quantified by using Quantity One Software (Bio-Rad, USA) and normalized to mRNA levels for β-actin.

Li Y., Tang Q., Wang P., Qin J., Wu H., Lin J, & Huang Z. (2017). Dynamic Changes of Mitochondrial Fusion and Fission in Brain Injury after Cardiac Arrest in Rats. BioMed Research International, 2017, 1948070.

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