MLKL Antibody [M19F23] | Primary Antibodies

Application: Reactivity: Human

Lane 1: Hela, Lane 2: HUVEC, Lane 3: HT-29

Usage Information

Dilution
WB1:1000 IHC1:400 IF1:200

Application
WB, IHC, IF

Reactivity
Human

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
54 kDa 115 kDa, 35, 54 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Datasheet & SDS

Biological Description

Specificity
MLKL Antibody [M19F23] detects endogenous levels of total MLKL protein.

Clone
M19F23

Synonym(s)
Mixed lineage kinase domain-like protein; hMLKL; MLKL

Background
MLKL, recognized as mixed lineage kinase domain-like protein, functions as the terminal effector of necroptosis, a regulated form of necrosis distinct from apoptosis, operating downstream of the receptor-interacting protein kinase 3 (RIPK3) within the necrosome complex to execute plasma membrane rupture. It features an N-terminal four-helix bundle (4HB) domain critical for membrane translocation and a pseudokinase domain that senses phosphorylation signals without catalytic activity. Upon RIPK3-mediated phosphorylation at key sites like T357/S358 in humans or S345 in mice, MLKL undergoes a conformational shift, oligomerizing into amyloid-like filaments that bind phosphatidylinositol phosphates on the inner plasma membrane leaflet, driving non-selective ion channel formation, osmotic imbalance, and rapid cell lysis. This mechanism integrates tightly with RIPK1/RIPK3 signaling initiated by death receptor ligands such as TNF-α under caspase-compromised conditions, where ubiquitylation at lysine 219 further enhances K63-linked chain formation to stabilize higher-order assemblies essential for cytotoxicity and pathogen clearance during infection. Activated MLKL not only terminates necroptosis but triggers concurrent inflammatory cascades by releasing damage-associated molecular patterns (DAMPs), activating the NLRP3 inflammasome for IL-1β maturation independently of gasdermin D, and amplifying chemokine production like IL-6 and MCP-1 to recruit immune effectors, positioning it as a dual regulator of cell death and innate immunity that researchers target to dissect sterile inflammation or microbial defense. MLKL maintains tissue homeostasis by curbing excessive necroptosis in barrier epithelia and neurons, with its deficiency mitigating low-grade sterile inflammation across skeletal muscle, adipose, and connective tissues during aging, while enabling survival advantages against viral challenges through heightened lymphocyte responses. Dysregulation drives pathology in inflammatory bowel disease, where p-MLKL exacerbates epithelial loss and cytokine storms, and neurodegenerative disorders like ALS or PD models, where MLKL knockout reduces neuroinflammation, motor deficits, and α-synuclein toxicity via lowered proinflammatory cytokines.

Background

MLKL, recognized as mixed lineage kinase domain-like protein, functions as the terminal effector of necroptosis, a regulated form of necrosis distinct from apoptosis, operating downstream of the receptor-interacting protein kinase 3 (RIPK3) within the necrosome complex to execute plasma membrane rupture. It features an N-terminal four-helix bundle (4HB) domain critical for membrane translocation and a pseudokinase domain that senses phosphorylation signals without catalytic activity. Upon RIPK3-mediated phosphorylation at key sites like T357/S358 in humans or S345 in mice, MLKL undergoes a conformational shift, oligomerizing into amyloid-like filaments that bind phosphatidylinositol phosphates on the inner plasma membrane leaflet, driving non-selective ion channel formation, osmotic imbalance, and rapid cell lysis. This mechanism integrates tightly with RIPK1/RIPK3 signaling initiated by death receptor ligands such as TNF-α under caspase-compromised conditions, where ubiquitylation at lysine 219 further enhances K63-linked chain formation to stabilize higher-order assemblies essential for cytotoxicity and pathogen clearance during infection. Activated MLKL not only terminates necroptosis but triggers concurrent inflammatory cascades by releasing damage-associated molecular patterns (DAMPs), activating the NLRP3 inflammasome for IL-1β maturation independently of gasdermin D, and amplifying chemokine production like IL-6 and MCP-1 to recruit immune effectors, positioning it as a dual regulator of cell death and innate immunity that researchers target to dissect sterile inflammation or microbial defense. MLKL maintains tissue homeostasis by curbing excessive necroptosis in barrier epithelia and neurons, with its deficiency mitigating low-grade sterile inflammation across skeletal muscle, adipose, and connective tissues during aging, while enabling survival advantages against viral challenges through heightened lymphocyte responses. Dysregulation drives pathology in inflammatory bowel disease, where p-MLKL exacerbates epithelial loss and cytokine storms, and neurodegenerative disorders like ALS or PD models, where MLKL knockout reduces neuroinflammation, motor deficits, and α-synuclein toxicity via lowered proinflammatory cytokines.

References
https://pubmed.ncbi.nlm.nih.gov/25288762/ https://pubmed.ncbi.nlm.nih.gov/34099649/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%