Western blot analysis was performed as described previously [29] (link), using the following antibodies: rabbit anti-MLKL, rabbit anti-RIPK3, rabbit anti-phospho MLKL, rabbit anti-phospho RIKP1, rabbit anti-phospho RIPK3, rabbit anti-TNFR1, rabbit anti-cIAP2 (all from Cell Signaling), mouse anti-RIPK1 (BD Bioscience), mouse anti-GAPDH (BioTrend), rabbit anti-TRAIL-R1 (Abcam), rabbit anti-TRAIL-R2 (Millipore), rabbit anti-FasR, mouse anti-cIAP1 (all from Santa Cruz), mouse anti-Caspase-8 (Enzo). For detection, secondary antibodies goat anti-mouse IgG and goat anti-rabbit IgG (Abcam) conjugated to horseradish peroxidase and enhanced chemiluminescence were used (Amersham Bioscience).
Rabbit anti phospho mlkl
Manufactured by Cell Signaling Technology
Sourced in United States
Rabbit anti-phospho MLKL is a primary antibody that specifically detects phosphorylated MLKL (mixed lineage kinase domain-like protein). MLKL is a key mediator of necroptosis, a form of programmed cell death. This antibody can be used to detect and monitor MLKL phosphorylation, a critical step in the necroptosis signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti phospho ripk3, by Cell Signaling Technology (2 mentions)
Mouse anti ripk1, by BD (2 mentions)
Rabbit anti ciap2, by Cell Signaling Technology (1 mentions)
Mouse anti gapdh, by Biotrend (1 mentions)
Mouse anti ciap1, by Santa Cruz Biotechnology (1 mentions)
Mouse anti caspase 8, by Enzo Life Sciences (1 mentions)
Goat anti rabbit igg, by Abcam (1 mentions)
Goat anti mouse igg, by Abcam (1 mentions)
Supersignal west pico plus chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Rabbit anti caspase 1 p10, by Santa Cruz Biotechnology (1 mentions)
Mouse anti caspase 1, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti cleaved caspase 1, by Cell Signaling Technology (1 mentions)
Rabbit anti gasdermin d, by Cell Signaling Technology (1 mentions)
Rabbit anti mouse cleaved caspase 8, by Cell Signaling Technology (1 mentions)
Mouse anti β actin, by Santa Cruz Biotechnology (1 mentions)
Goat anti mouse igg hrp, by Bio-Rad (1 mentions)
Chemidoc mp imager, by Bio-Rad (1 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (1 mentions)
Goat anti rabbit igg hrp, by Merck Group (1 mentions)
Rabbit anti ripk3, by ProSci (1 mentions)
3 protocols using rabbit anti phospho mlkl
1
Comprehensive Western Blot Analysis of Cell Death Regulators
2
Western Blot Analysis of Cell Apoptosis
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Cell lysates were obtained by lysing cells in 1% SDS lysis buffer containing 1% β-mercaptoethanol (Gibco). The lysates were immediately boiled for 10 min to minimize protein degradation. Supernatants were diluted in SDS buffer and boiled for 10 min prior to loading. Western blot analysis was performed using the following antibodies: rabbit anti-cleaved caspase-1 (89332S; Cell Signaling Technology), rabbit anti-gasdermin D (39754S; Cell Signaling Technology), rabbit anti-mouse cleaved caspase-8 (8592P; Cell Signaling Technology), mouse anti-RipK1 (610459; BD Biosciences), rabbit anti-RipK3 (2283; ProSci Inc), rabbit anti-phospho RipK3 (91702S; Cell Signaling Technology), rabbit anti-phospho MLKL (37333S; Cell Signaling Technology), rabbit anti-caspase-1 p10 (SC-514; Santa Cruz Biotechnology), mouse anti-caspase-1 (sc-56036; Santa Cruz Biotechnology), mouse anti-β-actin (SC-81178; Santa Cruz Biotechnology), goat anti-mouse IgG HRP (172-1011; Bio-Rad Laboratories Inc), and goat anti-rabbit IgG HRP (A6154; MilliporeSigma). The blots were subsequently detected by chemiluminescence with SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific Inc) on a ChemiDoc MP imager (Bio-Rad Laboratories Inc).
3
Western Blot Analysis of Protein Expression
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Western blot analysis was performed as described previously [25] (link) using the following antibodies: Mouse anti-STAT1 (Cell Signaling, Beverly, MA, USA), rabbit anti-phospho-STAT1 (Cell Signaling), mouse anti-IRF1 (Santa Cruz Biotechnologies, Santa Cruz, CA, USA), rabbit anti-MLKL (GeneTex, Inc., Irvine, CA, USA), rabbit anti-phospho-MLKL (Cell Signaling) mouse anti-GAPDH (HyTest, Turku, Finland), mouse anti-β-Actin (Sigma-Aldrich), mouse anti-Vinculin (Sigma-Aldrich). Goat anti-mouse IgG or goat anti-rabbit IgG conjugated to horseradish peroxidase (Santa Cruz Biotechnologies) and enhanced chemiluminescence (Amersham Bioscience, Freiburg, Germany) or infrared dye-labeled secondary antibodies and infrared imaging (Odyssey Imaging System, LI-COR Bioscience, Bad Homburg, Germany) were used for detection. Representative blots of at least two independent experiments are shown. Protein expressions of Western blots were quantified using ImageJ 1.52e and normalized to β-Actin protein expression.
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