At the end of the Langendorff RV-IR experiment, the RV was dissected free from the LV plus septum (LV+S) and weighed. The tissues were then preserved at −80 °C for subsequent immunoblotting. RV proteins were extracted from the various subcellular compartments using a cytoplasmic, mitochondrial and nuclear protein extraction kit (Cat #: MCN-002, ZmTech Scientifique; Montreal, QC, Canada). The following antibodies were used: anti-Drp 1 antibody (1:500; Cat #: ab56788, Abcam; Cambridge, MA, USA), anti-β-tubulin antibody (1:1000; Cat #: 32–2600, Invitrogen; Grand Island, NY, USA), anti-voltage-dependent anion channel (VDAC) antibody (1:1000; Cat #: 4866, Cell Signaling; Whitby, ON, Canada), anti-phospho-Drp 1 (serine 616) antibody (1:1000; Cat #: 3455S, Cell signaling; Whitby, ON, Canada), and anti-Fis 1 antibody (1:500; Cat #: 10956-1-AP, ProteinTech; Rosemont, IL, USA). β-tubulin and VDAC were used as cytoplasmic and mitochondrial loading controls, respectively.
Anti β tubulin antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Anti-β-tubulin antibody is a primary antibody used to detect and quantify the presence of the beta-tubulin protein in biological samples. It is a highly specific tool for researchers studying microtubule structure and dynamics in cells.
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Lab products found in correlation
Anti fis 1 antibody, by Proteintech (1 mentions)
Anti drp1 antibody, by Abcam (1 mentions)
In cell analyzer 2000, by GE Healthcare (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Anti gfap antibody, by Abcam (1 mentions)
2 protocols using anti β tubulin antibody
1
Subcellular Fractionation and Immunoblotting of Right Ventricular Proteins
2
Immunofluorescence Staining of Neurons
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The drug-treated neurons were fixed with 4% paraformaldehyde (Sigma–Aldrich) at room temperature. After three washes with PBS, the cells were permeabilized with blocking buffer (0.3% Triton X-100, 3% BSA, and 10% goat serum in PBS). The cells were then incubated with primary antibodies, including anti-MAP2 antibody (Novex, San Diego, CA, 1:500) or anti-β-Tubulin antibody (Invitrogen, CA, 1:500) and anti-GFAP antibody (Abcam, USA, 1:1000) at 4°C overnight, and followed with corresponding secondary antibodies for 1 h at room temperature in the dark. Finally, the cell nuclei were counterstained with 5 μg/ml of DAPI (Sigma–Aldrich) for 15 min. The samples were photographed using an InCell2000 Analyzer (GE Healthcare, USA).
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