Cell cultures were grown on poly-l-lysine-coated glass slides, fixed at room temperature for 10 min with 4% paraformaldehyde in PBS, permeabilized with 0.1% Triton X-100 for 5 min and then washed in PBS. Cells were blocked for 30 min in 2% horse serum in PBS and incubated with primary antibodies (mouse anti-ATPB (Abcam Cambridge, UK) and rabbit anti-HIF1 (Santa Cruz Santa Cruz CA, USA) diluted in blocking solution, for 1 h at 37°C and then with secondary antibodies Cy3- and Alexa488-conjugated (Molecular Probe Eugene OR, USA). After rinsing in PBS, cells were counterstained with 1 μg /ml Hoechst 33342 and examined with a Zeiss LSM 510 Confocal Laser Scanning Microscope. Fluorescence images were processed using ZEN 2009 (Carl Zeiss, Milan, Italy) and CorelDRAW image software.
Mouse anti atpb
Manufactured by Abcam
Sourced in United Kingdom
Mouse anti-ATPB is a primary antibody that recognizes the beta subunit of the mitochondrial ATP synthase complex. It is used for the detection and analysis of ATP synthase in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
Zen 2009, by Zeiss (1 mentions)
Lsm 510 confocal laser scanning microscope, by Zeiss (1 mentions)
Imagequant las 4000 imaging system, by GE Healthcare (1 mentions)
Clarity ecl, by Bio-Rad (1 mentions)
Peroxidase conjugated goat anti mouse igg, by Bio-Rad (1 mentions)
Oxphos human wb antibody cocktail, by Abcam (1 mentions)
Mouse anti mtco1, by Abcam (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Alexa fluor 546 anti rabbit, by Thermo Fisher Scientific (1 mentions)
Chicken anti map2, by Abcam (1 mentions)
Rabbit anti tom20, by Proteintech (1 mentions)
Anti mouse alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Fluoroshield with dapi, by Merck Group (1 mentions)
Goat serum, by Merck Group (1 mentions)
3 protocols using mouse anti atpb
1
Immunostaining of HIF1 and ATPB in Cells
2
Quantifying Mitochondrial Protein Levels
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Protein samples were separated by polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride (PVDF) membranes (Merck Millipore). Blots were blocked with 5% nonfat dry milk in tris-buffered saline containing 0.05% Tween-20 (TBST) for 1 h and then incubated with primary antibodies. The following antibodies were used: mouse anti-MTCO1 (1:2,000, Abcam, Cambridge, UK), mouse anti-complex IV subunit (COX IV; 1:2,000, Abcam), mouse anti-ATPB (1:2,000, Abcam), Total Oxidative Phosphorylation (OXPHOS) Human WB Antibody Cocktail (1:200, Abcam), mouse anti-VDAC1/Porin (1:1,000, Abcam). Peroxidase-conjugated goat antimouse IgG (1:10,000, Bio-Rad) and antirabbit IgG (1:5,000, Bio-Rad) were used as secondary antibodies. To control for the amount of protein loaded, we used a mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH)-GAPDH-loading control antibody (1:10,000, Abcam). Immunoreactive proteins were visualized using an enhanced chemiluminescence detection system (Clarity ECL, Bio-Rad), and bands were detected with an ImageQuant LAS 4000 Imaging System (GE Healthcare Life Sciences, Barcelona, Spain).
3
Immunofluorescent Staining of Cultured Cells
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Cells were fixed with 4% paraformaldehyde (ThermoFisher, 50980487) for 15 min at 37 °C. Coverslips were incubated in blocking solution (5% goat serum (Sigma, G9023) and 0.3% Triton-X100 (Acros Organics, 215682500) in PBS) for 60 min. Coverslips were then incubated in primary antibody solution (1:1000 Mouse Anti-ATPB (Abcam, ab14730), 1:200 Rabbit Anti-TOM20 (ProteinTech, 11802–1-AP), 1:1000 Chicken Anti-MAP2 (Abcam, ab5392), 1% BSA (Sigma, A9647) and 0.3% Triton-X100 in PBS) at 4 °C overnight. After primary incubation, coverslips were washed 3 × in PBS and incubated in secondary antibody solution (1:200 Anti-Mouse Alexa Fluor 488 (Invitrogen, A11029), 1:200 Anti-Rabbit Alexa Fluor 546 (Invitrogen, A10040), 1:200 Anti-Chicken Alexa Fluor 647 (Invitrogen, A21449), 1% BSA (Sigma, A9647) and 0.3% Triton-X100 in PBS) for 60 min. Following secondary incubation, coverslips were washed 3 × with PBS and mounted on glass slides using Fluoroshield with DAPI (Sigma, F6057).
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