Preparation and Characterization of Antibodies for Apoptosis Regulators

Rabbit polyclonal antibodies against caspase-1, NLRP3 and ASC were made in house and were described previously56 (link)57 (link)58 (link). T7̇Tag monoclonal antibody HRP conjugate (Catalogue No. 69048) was from Novagen. Monoclonal anti-β-actin (Catalogue No. A-5316) was from Sigma Aldrich. Anti-DFNA5 (Catalogue No. sc-79233), anti-caspase-3 (Catalogue No. sc-7148), anti-GSDMD (Catalogue No. sc-393656) and anti-Bax (Catalogue No. sc-930) were from Santa Cruz. Anti- Na,K-ATPase polyclonal antibody (Catalogue No. 3010) was from Cell Signaling Technology. zVAD-fmk (Catalogue No. A1902) was obtained from ApexBio. GSK'872 (Cat No. AOB4887) was obtained from Aobious. CytoTox96 LDH-release kit (Cat No. G1780) was from Promega. TALON metal affinity resin (Catalogue No. 635502) and In-Fusion HD Cloning Plus (Catalogue No. 638910) were obtained from Takara Clontech. All antibodies were used at 1/1,000–1/2,000 dilutions for western blot analyses.

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Rogers C., Fernandes-Alnemri T., Mayes L., Alnemri D., Cingolani G, & Alnemri E.S. (2017). Cleavage of DFNA5 by caspase-3 during apoptosis mediates progression to secondary necrotic/pyroptotic cell death. Nature Communications, 8, 14128.

Publication 2017

Monoclonal anti-β-actin anti-caspase-3 anti-GSDMD anti-Bax zVAD-fmk GSK'872 CytoTox96 LDH-release kit TALON metal affinity resin In-Fusion HD Cloning Plus

Actin Antibodies Antibody Atpase Caspase 1 Caspase 3 Dfna5 Dilutions Metal Monoclonal antibody Promega Rabbit Resin Talon Western blot analyses Zvad fmk

Corresponding Organization :

Other organizations : Thomas Jefferson University, Sidney Kimmel Cancer Center, Pennsylvania State University

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Variable analysis

independent variables

Rabbit polyclonal antibodies against caspase-1
Rabbit polyclonal antibodies against NLRP3
Rabbit polyclonal antibodies against ASC
T7 Tag monoclonal antibody HRP conjugate
Monoclonal anti-β-actin
Anti-DFNA5
Anti-caspase-3
Anti-GSDMD
Anti-Bax
Anti-Na,K-ATPase polyclonal antibody
ZVAD-fmk
GSK'872

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

positive controls

Not specified

negative controls

Not specified

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