Cell culture supernatant were precipitated with methanol and chloroform, as previously described, and resuspended cell extracts lysed in boiling lysis buffer [66 mM tris-Cl (pH 7.4), 2% SDS, 10 mM dithiothreitol, and NuPage LDS sample buffer; Thermo Fisher Scientific). Proteins were separated on 14% polyacrylamide gels and transferred onto nitrocellulose membrane using Trans-Blot Turbo (Bio-Rad). Antibodies for immunoblot were against GSDMD (1:3000; EPR19828, Abcam), caspase-1 p20 (1:1000; casper-1, AdipoGen), full-length caspase-8 (1:1000; 4927, Cell Signaling), cleaved caspase-8 (1:1000; 9429, Cell Signaling), caspase-3 (1:1000; 9662, Cell Signaling), pro–IL-1β (1:1000; AF-401-NA, R&D), IL-18 (1:1000; 5180r-100, BioVision), RIPK1 (1:1000; D94C12, Cell Signaling), c-FLIP (1:1000; Dave-2, AdipoGen), and alpha-tubulin (1:5000; DM1A, Abcam).
Dave 2
Manufactured by Adipogen
Sourced in United States
Dave-2 is a laboratory device designed for the efficient purification and isolation of proteins. It utilizes a proprietary chromatography technique to separate and concentrate target proteins from complex biological samples. The core function of Dave-2 is to provide a reliable and consistent method for the purification of proteins for further analysis and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Af 401 na, by R&D Systems (1 mentions)
Epr19828, by Abcam (1 mentions)
Trans blot turbo, by Bio-Rad (1 mentions)
Nupage lds sample buffer, by Thermo Fisher Scientific (1 mentions)
Casper 1, by Adipogen (1 mentions)
Anti mbp, by New England Biolabs (1 mentions)
Anti dr5, by Cell Signaling Technology (1 mentions)
Anti dr4, by ProSci (1 mentions)
Killer trail, by Alexis Biochemicals (1 mentions)
Anti fadd 556402, by BD (1 mentions)
β actin, by Merck Group (1 mentions)
2 protocols using dave 2
1
Immunoblotting of Inflammasome Proteins
2
Evaluation of cFLIP-targeting Molecules
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
KillerTRAIL (human recombinant) was purchased from Alexis Biochemicals. The nine most highly selective molecules targeting cFLIP were obtained from NCI–DSCB (National Cancer Institute-Drug Synthesis and Chemistry Branch, Rockville, MD, USA). For Western blotting (WB) experiments, we used the following antibodies: anti-FLIP antibodies DAVE2 and NF6 (Adipogen, San Diego, CA, USA), caspase-3 (8G10; OZYME), PARP (Asp214, 19F4; OZYME, Saint-Cyr-l’École, France), caspase-8 (1C12; OZYME), anti-MBP (New England Biolabs, Ipswich, MA, USA) and anti-His (C-ter) (Invitrogen, Carlsbad, CA, USA). Anti-His (ab81663) was used to immunoprecipitate the DISC complex, and the following antibodies were used for WB analysis: anti-Flip (DAVE II, Adipogen), anti-FADD (556402, BD, San Jose CA, USA), anti-casp8 (5F7, EnzoLife, Farmingdale, NY, USA), anti-DR4 (1139, ProSci, Poway, CA, USA) and anti-DR5 (3696, Cell Signaling, Danvers, MA, USA). Anti-rabbit and anti-mouse HRP-linked secondary antibodies (Santa Cruz Biotechnology, Dallas, TX, USA) and β-actin (Sigma Aldrich, St. Louis, MO, USA) were also used.
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!