Antibodies and Reagents for MLKL and RIPK3

Primary antibodies used in this study were: rat anti-MLKL (clone 3H1, produced in-house; 1:1000 dilution; available as MABC604, EMD Millipore, Billerica, MA, USA), rat anti-human MLKL pseudokinase domain (clone 7G2, produced in-house^{32 (link)}; 1:2000 dilution; soon available from EMD Millipore as MABC1636), rabbit anti-human MLKL phospho-S358 (AB187091, Abcam; 1:2000), mouse anti-Actin (C4) HRP (sc-47778 HRP, Santa Cruz Biotechnology; 1:10000), rat anti-human RIPK3 (clone 1H2, produced in-house^{11 (link)}; 1:1000; available as MABC1640, EMD Millipore, Billerica, MA, USA), rabbit anti-human RIPK3 phospho-S227 (D6W2T, CST; 1:2000). Recombinant hTNF-Fc^{62 (link)} was produced in-house, while the Smac mimetic, Compound A^{63 (link)} and the pan-caspase inhibitor, IDN-6556/Emricasan, were provided by Tetralogic Pharmaceuticals. The RIPK3 inhibitors, GSK′843 (ref. ^{54 (link)}) and BMS Compound 10 (ref. ^{48 (link)}), were kindly provided by Anaxis Pty Ltd (Australia).

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Meng Y., Davies K.A., Fitzgibbon C., Young S.N., Garnish S.E., Horne C.R., Luo C., Garnier J.M., Liang L.Y., Cowan A.D., Samson A.L., Lessene G., Sandow J.J., Czabotar P.E, & Murphy J.M. (2021). Human RIPK3 maintains MLKL in an inactive conformation prior to cell death by necroptosis. Nature Communications, 12, 6783.

Publication 2021

MABC604 AB187091 (sc-47778 HRP, MABC1640

Actin Antibodies Caspase inhibitor Clone Dilution Emricasan Human Idn 6556 Inhibitors Mouse Pharmaceuticals Rabbit Ripk3

Corresponding Organization :

Other organizations : Walter and Eliza Hall Institute of Medical Research, University of Melbourne

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Variable analysis

independent variables

Recombinant hTNF-Fc
Smac mimetic, Compound A
Pan-caspase inhibitor, IDN-6556/Emricasan
RIPK3 inhibitors, GSK'843 and BMS Compound 10

dependent variables

MLKL phospho-S358
RIPK3
RIPK3 phospho-S227

control variables

Actin

controls

Positive controls: Not explicitly mentioned.
Negative controls: Not explicitly mentioned.

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