Rabbit anti opa1

Manufactured by Abcam

Sourced in United States

Rabbit anti-OPA1 is a primary antibody that specifically recognizes the OPA1 (Optic Atrophy 1) protein. OPA1 is a dynamin-related GTPase that plays a key role in mitochondrial inner membrane fusion and cristae organization. This antibody can be used for applications such as Western blotting, immunoprecipitation, and immunofluorescence.

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Anti-OPA1 (17 citations) Rabbit anti- (5 citations) Polyclonal rabbit anti-OPA1 (3 citations) Rabbit anti-OPA1 antibody (2 citations)

3 protocols using rabbit anti opa1

Mitochondrial Dynamics Analysis in Human EPCs

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Human EPCs were lysed with Pro-prep (iNtRON biotechnology, Korea). Whole cell lysates (25 µg) were separated by either 8% or 15% SDS-PAGE, after which membranes were blocked with 5% skim milk for 1 h at 25℃. Membranes were probed with specific antibodies, and proteins of interest were visualized with HRP-conjugated secondary antibodies (Thermo Fisher Scientific Inc., MA, USA) and Luminata Crescendo HRP substrates (Millipore, MA, USA). Primary antibodies against mouse anti-HIF-1A (1:1000), mouse anti-DLP1 (1:1000, all above from BD Biosciences, NJ, USA), rabbit anti-phospho-DRP1 (Ser616) (1:1000), rabbit anti-phospho-DRP1 (Ser637) (1:1000, Cell Signaling Technology, Inc., MA, USA), rabbit anti-Fis1 (1:1000), rabbit anti-OPA1 (1:1000, all above Abcam, Cambridge, UK), rabbit anti-MFN1 (1:1000), and mouse anti-beta-actin (1:2500, all above Santa Cruz Biotechnology, CA, USA) were used.

Kim D.Y., Jung S.Y., Kim Y.J., Kang S., Park J.H., Ji S.T., Jang W.B., Lamichane S., Lamichane B.D., Chae Y.C., Lee D., Chung J.S, & Kwon S.M. (2018). Hypoxia-dependent mitochondrial fission regulates endothelial progenitor cell migration, invasion, and tube formation. The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology, 22(2), 203-213.

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Mitochondrial Dynamics in Cybrids

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Mitochondrial fraction isolated from cybrid cells were suspended in buffer (150mM KCl, 5mM HEPES, 2mM K₂HPO₄, 5mM glutamate, 5mM malate, 150mM potassium thiocyanate, pH 7.2). Mitochondrial fraction and cell lysate were subjected to immunoblotting. Mouse anti-Mfn2 (1:2000, Sigma), rabbit anti-Drp1 (1:3000, Thermo scientific), rabbit anti-Opa1 (1:5000, Abcam), rabbit anti-Fis1 (1:3000, Abcam), mouse anti-Hsp60 (1:5000, Enzo), rabbit anti-phospho-ERK1/2, mouse anti-ERK1/2 (1:2000, Cell signaling), rabbit anti-PGC1α (1:2000, Santa Cruz) and mouse anti-β-actin (1:8000, Sigma) were used as primary antibodies. Binding sites of primary antibody were visualized with horseradish peroxidase-conjugated anti-rabbit IgG antibody (1:5000, life technology) or anti-mouse IgG antibody (1:5000, life technology) followed by the addition of enhanced chemiluminescence (ECL) substrate (GE Healthcare). Relative quantification of optical density for immune reactive bands was performed using NIH Image J software.

Gan X., Wu L., Huang S., Zhong C., Li G., Yu H., Swerdlow R.H., Chen J.X, & Yan S.S. (2014). Oxidative stress-mediated activation of extracellular signal-regulated kinase contributes to mild cognitive impairment-related mitochondrial dysfunction. Free radical biology & medicine, 75, 230-240.

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Protein extraction and Western blot

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Total cell lysates were generated in RIPA buffer (1% Nonidet P40, 0.5% sodium deoxycholate, 0.1% SDS in PBS). Protein was quantified using the Pierce BCA Protein Assay Kit (Thermo Scientific) according to manufacturer’s instructions. Westerns were performed as described [30] (link). PVDF was immunoblotted with antibodies against HTRA2 (R&D Systems), HSP90 or β-actin (Santa Cruz), OPA1 (mouse anti-OPA1 from BD Pharminogen, rabbit anti-OPA1 from Abcam), VDAC, MFN2, LC3β, P62 or PHB2 (Cell Signaling). ImageJ was used to quantify band intensity to calculate OPA1 processed form ratios from at least three animals per genotype.

Patterson V.L., Zullo A.J., Koenig C., Stoessel S., Jo H., Liu X., Han J., Choi M., DeWan A.T., Thomas J.L., Kuan C.Y, & Hoh J. (2014). Neural-Specific Deletion of Htra2 Causes Cerebellar Neurodegeneration and Defective Processing of Mitochondrial OPA1. PLoS ONE, 9(12), e115789.

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