For immunoprecipitation and Western blotting [50 (link)], mouse anti-PARP-1 polyclonal antibody, mouse pan-Acetylation monoclonal antibody and mouse β-Actin monoclonal antibody was purchased from Proteintech (Proteintech Group, Chi cago, IL, USA). Mouse anti-PAR-monoclonal antibody was purchased from Trevigen (Trevigen Inc., Gaithersburg, Maryland, USA). Rabbit anti-sirtuin 3 (SIRT3) polyclonal antibody were purchased from Cell Signaling Technology (Beverly, MA, USA) and Proteintech (Proteintech Group, Chi cago, IL, USA). Nuclear proteins were extracted with a commercially available Nuclear and Cytoplasm Extraction kit (Active Motif, Carlsbad, CA, USA) according to the manufacturer’s recommendations. Western blot analyses were performed as previously described [19 (link), 51 (link)] and β-Actin was used as a loading control. For co-IP, total proteins (400μg) incubated with 1μg anti-PARP-1 antibody for overnight (mouse normal IgG was used as a control), or nuclear proteins (200-300μg) incubated with 1 μg anti-SIRT3 antibody for overnight (rabbit normal IgG was used as a control), followed by 4h incubation with protein A/G PLUS-Agarose (Santa Cruz, CA, USA) at 4°C. The co-IP proteins were detected by Western blot.
Mouse β actin monoclonal antibody
Manufactured by Proteintech
Sourced in United States
The Mouse β-Actin monoclonal antibody is a laboratory reagent used to detect the presence and quantity of the β-Actin protein in mouse biological samples. It is a specific antibody that binds to the β-Actin protein, allowing researchers to quantify or visualize its expression in various experimental contexts.
Automatically generated - may contain errors
Lab products found in correlation
Nuclear and cytoplasm extraction kit, by Active Motif (1 mentions)
Mouse monoclonal anti par antibody, by Bio-Techne (1 mentions)
Protein a g plus agarose, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti bcl 2 antibody, by Cell Signaling Technology (1 mentions)
Rabbit anti bax antibody, by Proteintech (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Polyvinylidene difluoride membrane, by Bio-Rad (1 mentions)
2 protocols using mouse β actin monoclonal antibody
1
Immunoprecipitation and Western Blot Analysis
2
Western Blot Analysis of Liver IRI
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteins were extracted from liver IRI tissues and cultured with BMDM using RIPA Lysis Buffer (Beyotime, Shanghai, China) for Western blotting.
Protein concentrations were calculated using the BCA Protein Assay Kit (Beyotime, Shanghai, China). Equal amounts of protein (50 μg) were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride membranes (Bio-Rad, Hercules, CA, USA). Membranes were blocked in 5% nonfat milk and then incubated overnight at 4 ℃ with the following primary antibodies: rabbit anti-Bax antibody (1∶2000; Cat. #50599-2-lg, Proteintech), rabbit anti-Bcl2 antibody (1∶2000; Cat. #4223T, Cell Signaling Technology, Massachusetts, USA, ), mouse anti-TLR4 antibody (1∶2000; Cat. #66350-1-1g, Proteintech), rabbit anti-MyD88 antibody(1∶2000; Cat. #23230-1-AP, Proteintech), rabbit anti-p65 antibody(1∶2000, Cat. #80979-1-RR, Proteintech), rabbit anti-p-p65 antibody(1∶2000; Cat. #82335-1-RR, Proteintech), and mouse β-actin monoclonal antibody (1∶5000; Cat. #66009-1-1g, Proteintech). βactin was used as a control. The next day, membranes were incubated with peroxidase-conjugated goat antirabbit or goat anti-mouse IgG for 1 h at room temperature and subjected to substrate development.
Protein concentrations were calculated using the BCA Protein Assay Kit (Beyotime, Shanghai, China). Equal amounts of protein (50 μg) were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride membranes (Bio-Rad, Hercules, CA, USA). Membranes were blocked in 5% nonfat milk and then incubated overnight at 4 ℃ with the following primary antibodies: rabbit anti-Bax antibody (1∶2000; Cat. #50599-2-lg, Proteintech), rabbit anti-Bcl2 antibody (1∶2000; Cat. #4223T, Cell Signaling Technology, Massachusetts, USA, ), mouse anti-TLR4 antibody (1∶2000; Cat. #66350-1-1g, Proteintech), rabbit anti-MyD88 antibody(1∶2000; Cat. #23230-1-AP, Proteintech), rabbit anti-p65 antibody(1∶2000, Cat. #80979-1-RR, Proteintech), rabbit anti-p-p65 antibody(1∶2000; Cat. #82335-1-RR, Proteintech), and mouse β-actin monoclonal antibody (1∶5000; Cat. #66009-1-1g, Proteintech). βactin was used as a control. The next day, membranes were incubated with peroxidase-conjugated goat antirabbit or goat anti-mouse IgG for 1 h at room temperature and subjected to substrate development.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!