DIE cells were harvested by trypsinization and lysed with RIPA buffer. Total protein concentrations were determined using a Pierce BCA protein assay kit (Fischer Scientific). 20ug protein was denatured using 4x Laemmli sample buffer (Bio-Rad) with 10% BME (Sigma), and boiled for 5 minutes. Samples were run on a 15% SDS-polyacrylamide gel and transferred to a PVDF membrane (Merck). Membranes were blocked for an hour using 5% BSA in TBST, and were subsequently incubated overnight with anti-DUX4 antibody [E5-5] (Abcam, ab124699) in blocking solution (5% BSA in TBST), at 4°C. Membranes were than incubated for an hour with secondary goat anti-rabbit-HRP antibody (Santa Cruz, sc-2004), and primary rabbit mAb β-Actin HRP conjugated antibody (Cell signaling, 5125s) in blocking buffer. Chemiluminescent signal was detected using GE ImageQuant LAS 4000 imager, upon admission of Pierce ECL Plus Western Blotting substrate (Fischer Scientific).
Anti dux4 antibody e5 5
Manufactured by Abcam
The Anti-DUX4 antibody [E5-5] is a research-use only product. It is a monoclonal antibody that recognizes the DUX4 protein. DUX4 is a transcription factor involved in muscle development and facioscapulohumeral muscular dystrophy (FSHD).
Automatically generated - may contain errors
Lab products found in correlation
Goat anti rabbit hrp secondary antibody, by Santa Cruz Biotechnology (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Laemmli sample buffer, by Bio-Rad (2 mentions)
Pierce ecl plus western blotting substrate, by Thermo Fisher Scientific (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
β actin hrp conjugated antibody, by Cell Signaling Technology (2 mentions)
Imagequant las 4000 imager, by Thermo Fisher Scientific (2 mentions)
2 protocols using anti dux4 antibody e5 5
1
Western Blot Analysis of DUX4 Protein
2
Western Blot Analysis of DUX4 Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DIE cells were harvested by trypsinization and lysed with RIPA buffer. Total protein concentrations were determined using a Pierce BCA protein assay kit (Fischer Scientific). 20ug protein was denatured using 4x Laemmli sample buffer (Bio-rad) with 10% BME (Sigma), and boiled for 5 minutes. Samples were run on a 15% SDSpolyacrylamide gel and transferred to a PVDF membrane (Merck). Membranes were blocked for an hour using 5% BSA in TBST, and were subsequently incubated overnight with anti-DUX4 antibody [E5-5] (Abcam, ab124699) in blocking solution (5% BSA in TBST), at 4°C. Membranes were than incubated for an hour with Secondary goat anti-rabbit-HRP antibody (Santa Cruz, sc-2004), and primary rabbit mAb β-Actin HRP conjugated antibody (Cell signaling, 5125s) in blocking buffer. Chemiluminescent signal was detected using GE ImageQuant LAS 4000 imager, upon admission of Pierce ECL Plus Western Blotting substrate (Fischer Scientific).
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!