RGCs were loaded with radio immunoprecipitation assay buffer. Lysates were collected by centrifugation at 12 000 rpm for 5 min at 4 °C. Proteins were separated by SDS-PAGE and transferred to polyvinylidene difluoride membranes for western blotting. Membranes were blocked for 1 h at RT in 5% nonfat dry milk, then incubated with polyclonal rabbit anti-parkin (1 : 1000; Abcam, USA), monoclonal rabbit anti-Bax (1 : 1000; Abcam), polyclonal rabbit anti-Bcl-2 (1 : 500; Abcam), monoclonal rabbit anti-LC3 (1 : 2000; Abcam), polyclonal rabbit anti-LAMP1 (1 : 1000; Abcam), polyclonal rabbit anti-optineurin (1 : 200; Abcam) and polyclonal rabbit anti-GAPDH (1 : 2000; Weiao Biotechnology Co, Shanghai, China) in primary antibody dilution (Weiao Biotechnology Co) at 4 °C overnight. After washing with TBST, membranes were incubated with secondary antibodies (1 : 2000; Jackson). Blots were visualized using enhanced chemiluminescence reagents (Weiao Biotechnology Co). Chemiluminescent images were captured on X-ray film using a developing and fixing solution in a dark room and at last analyzed with Image J (National Institutes of Health, USA).
Monoclonal rabbit anti bax
Manufactured by Abcam
Sourced in United States, China
Monoclonal rabbit anti-Bax is a primary antibody that specifically binds to the Bax protein. Bax is a pro-apoptotic member of the Bcl-2 protein family and plays a key role in the initiation of the mitochondrial apoptosis pathway.
Automatically generated - may contain errors
Lab products found in correlation
Polyclonal rabbit anti bcl 2, by Abcam (3 mentions)
Polyclonal rabbit anti parkin, by Abcam (3 mentions)
Polyclonal rabbit anti lamp1, by Abcam (2 mentions)
Monoclonal rabbit anti lc3, by Abcam (2 mentions)
Polyclonal rabbit anti optineurin, by Abcam (2 mentions)
Peroxidase conjugated goat anti rabbit igg, by Jackson ImmunoResearch (2 mentions)
Ripa buffer, by Beyotime (2 mentions)
Polyclonal rabbit anti opa1, by Abcam (2 mentions)
Polyclonal rabbit anti gapdh, by Yesen (2 mentions)
Supersignal west femto substrate trial kit, by Thermo Fisher Scientific (2 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Rabbit anti gfap polyclonal antibody, by Abcam (1 mentions)
Image station 4000mm pro, by Carestream (1 mentions)
3 protocols using monoclonal rabbit anti bax
1
Western Blot Analysis of Neurodegeneration Markers
2
Retinal ganglion cell protein analysis
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Retinas (n = 3 per group) were lysed in RIPA buffer (Beyotime, China) and ultrasonically smashed on ice to get homogenized solutions. RGCs (n = 3 per group) were mixed with RIPA buffer. Equal amounts of protein were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the electrotransferred onto polyvinylidene difluoride membranes (Millipore, Billerica, MA, United States). After blocking with 5% non-fat dry milk at room temperature for 1 h, the membranes were incubated with polyclonal rabbit anti-OPA1 (1:1000; Abcam), polyclonal rabbit anti-GFAP antibody (1:10000; Abcam), polyclonal rabbit anti-Parkin (1:1000; Abcam), polyclonal rabbit anti-Bcl-2 (1:500; Abcam), monoclonal rabbit anti-Bax (1:1,000; Abcam), monoclonal rabbit anti-LC3 (1:2000; Abcam), polyclonal rabbit anti-LAMP1 (1:1000; Abcam) and polyclonal rabbit anti-GAPDH (1:2000; Yesen, China) in primary antibody dilution (Beyotime, China) overnight at 4°C. After the membranes were washed several times, secondary antibody peroxidase-conjugated goat anti-rabbit IgG (1:5000; Jackson) was applied. Proteins were visualized by a Kodak Image Station 4000MM PRO (Carestream, Rochester, NY, United States) using chemiluminescence detection (SuperSignal West Femto Substrate Trial Kit, Thermo Fisher). Band intensity was analyzed with Image J (National Institutes of Health).
3
Protein Expression Analysis of Retinal Ganglion Cells
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RGCs (n = 4 per group) were mixed with RIPA buffer (Beyotime, Shanghai, China). Each sample (10 μg) was separated with polyacrylamide gel electrophoresis (PAGE) and electrotransferred on polyvinylidene difluoride (PVDF) membranes. Membranes were blocked with 5% nonfat dry milk at room temperature for 1 h, incubated with polyclonal rabbit anti-parkin (1:1,000; Abcam), polyclonal rabbit anti- optineurin (1:200; Abcam), polyclonal rabbit anti- Bcl-2 (1:500; Abcam), monoclonal rabbit anti- Bax (1:1,000; Abcam), polyclonal rabbit anti-OPA1 (1:1,000; Abcam), and polyclonal rabbit anti-GAPDH (1:2000; Yesen, Shanghai, China) in primary antibody dilution (Beyotime) at 4 °C overnight. The membranes were rinsed with 1X Tris-buffered saline/Tween 20 (TBST; Worthington) several times, incubated with peroxidase-conjugated goat anti-rabbit IgG (1:5,000; Jackson Laboratories, West Grove, PA), and then developed using chemiluminescence detection (SuperSignal™ West Femto Substrate Trial Kit, Thermo Fisher, Waltham, MA). Chemiluminescent images were captured using a Kodak Image Station 4000MM PRO (Carestream, Rochester, NY) and analyzed with Image J (National Institutes of Health).
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