The total protein extracts were prepared as described previously.29 (link) In brief, cells were lysed in cold radioimmunoprecipitation assay buffer with protease and phosphatase inhibitor cocktail (Roche). Equal amounts of protein samples were separated on SDS-PAGE, and then transferred onto nitrocellulose membranes (Millipore, Billerica, MA, USA). Then, membranes were probed with anti-PGC-1α (1:2,000; Santa Cruz Biotechnology, Dallas, TX, USA), anti-TFAM (1:16,000; Novus Bio, Littleton, CO, USA), anti-NRF1 (1:1,000; Santa Cruz Biotechnology), anti-SIRT1 (1:2,000; Sigma-Aldrich, St Louis, MO, USA), anti-cyto c (1:2,000; eBioscience, San Diego, CA, USA), anti-DIABLO (1:1,000; Cell Signaling Technologies, Danvers, MA, USA), and anti-GAPDH antibodies (1:1,000; Cell Signaling Technologies). The blots were then analyzed by chemiluminescence detection (ECL, Amersham, GE Healthcare Life Sciences, Chicago, IL, USA).
Anti diablo
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-DIABLO is a primary antibody that recognizes the DIABLO (also known as Smac) protein. DIABLO is a mitochondrial protein that plays a role in the induction of apoptosis. The antibody can be used to detect DIABLO expression in various cell and tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Nitrocellulose membrane, by Merck Group (1 mentions)
Anti tfam, by Novus Biologicals (1 mentions)
Anti nrf1, by Santa Cruz Biotechnology (1 mentions)
Anti sirt1, by Merck Group (1 mentions)
Anti cyto c, by Thermo Fisher Scientific (1 mentions)
Anti pgc 1α, by Santa Cruz Biotechnology (1 mentions)
Anti gapdh antibody, by Cell Signaling Technology (1 mentions)
Enhanced chemi luminescence (ecl), by GE Healthcare (1 mentions)
Protease and phosphatase inhibitor cocktail, by Roche (1 mentions)
Image lab 6, by Bio-Rad (1 mentions)
Anti mcl 1, by Cell Signaling Technology (1 mentions)
Chemidoc xrs system, by Bio-Rad (1 mentions)
2 protocols using anti diablo
1
Protein Expression Analysis Protocol
2
Western Blot Analysis of Apoptotic Regulators
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
T cells were lysed and suspended in Laemmli buffer. SDS-PAGE was performed under non-reducing conditions, and proteins were transferred to a 0.2-μm pore size polyvinylidene difluoride (PDFV) membrane (Bio-Rad, Hercules, CA, USA). Western blot was made using the following antibodies: anti-Mcl-1 (dilution 1:1,000; Cell Signaling Technology, Danvers, MA, USA), anti-DIABLO (dilution 1:1,000; Cell Signaling Technology), anti-glyceraldehyde 3-phospate dehydrogenase (GAPDH, dilution 1:1,000; GeneTex Biotechnology, Irvine, CA, USA), and anti-mouse- and anti-rabbit horseradish peroxidase-labeled IgG (both used 1:2,000; R&D Systems, Minneapolis, MN, USA). Protein bands were detected by incubation with enhanced chemiluminescence reagent (Thermo Scientific, Pierce Biotech., Rockford, IL, USA) and visualized with the Imaging System from Bio-Rad (ChemiDoc™ XRS+ System). Band densities were analyzed by densitometry using ImageLab 6.0.1 software provided by Bio-Rad. Each sample was normalized using GAPDH as a loading control.
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!