Rabbit anti opa1

Manufactured by Cell Signaling Technology

Sourced in United States

Rabbit anti-OPA1 is a primary antibody that specifically binds to the OPA1 protein. OPA1 is a mitochondrial protein involved in the regulation of mitochondrial dynamics, including mitochondrial fusion. This antibody can be used to detect and study the expression and localization of OPA1 in various cell and tissue samples.

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Close spelling variants of this product

Anti-OPA1 (13 citations) Rabbit monoclonal anti-Opa1 (2 citations)

2 protocols using rabbit anti opa1

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 Proteins

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Specific proteins expressed in the mouse brain and neuron cell cultures were determined by western blotting using specific antibodies. Proteins (40 µg) were separated on 10% or 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels and then transferred onto a nitrocellulose membrane. Nonspecific binding sites were blocked using 5% fat-free dried milk in 1 × TBST at 37 °C for 1 h. The membrane was incubated at 4 °C overnight with individual primary antibodies, including rabbit anti-OPA1 (Cell Signaling Technology, USA), rabbit anti-MFN1 (Proteintech, USA), mouse anti-MFN2 (Abcam, USA), rabbit anti-cytochrome c (Cell Signaling Technology, USA), rabbit anti-cleaved caspase 3 (Cell Signaling Technology, USA), mouse anti-GAPDH (Proteintech, USA), rabbit anti-β-actin (Proteintech, USA), and rabbit anti-VDAC (Cell Signaling Technology, USA). After washing with TBST, the membrane was incubated with secondary antibodies, goat anti-mouse IgG or anti-rabbit IgG conjugated to horseradish peroxidase (1:10,000). The blots were visualized using a Chemiluminescent Imaging System (Tanon Science and Technology).

Wu W., Zhao D., Shah S.Z., Zhang X., Lai M., Yang D., Wu X., Guan Z., Li J., Zhao H., Li W., Gao H., Zhou X., Qiao J, & Yang L. (2019). OPA1 overexpression ameliorates mitochondrial cristae remodeling, mitochondrial dysfunction, and neuronal apoptosis in prion diseases. Cell Death & Disease, 10(10), 710.

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