MLKL Antibody [H6A20]

Catalog No.: F4607

Application: Reactivity: Mouse

Lane 1: NIH/3T3, Lane 2: Neuro-2a

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代表番号: 045-509-1970|電子メール：sales@selleck.co.jp

使用情報

Dilution
WB1:1000 IP1:50

Application
WB, IP

Source
Rabbit Monoclonal Antibody

Reactivity
Mouse

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
54 kDa

Datasheet & SDS

生物学的記述

Specificity
MLKL Antibody [H6A20] detects endogenous levels of total MLKL protein.

Clone
H6A20

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

Background
MLKL (Mixed Lineage Kinase Domain-Like protein) is a pseudokinase of the protein kinase superfamily that acts as the terminal effector of necroptosis, a form of regulated necrotic cell death triggered by TNF or TLR ligands when caspases are inhibited. MLKL consists of an N-terminal four-helix bundle (4HB) executioner domain (α1–α4 helices that form membrane-disrupting pores), a central two-helix “brace” region connecting domains, and a C-terminal pseudokinase domain (PsKD) that, despite lacking catalytic activity, binds nucleotides and regulates activation. Key residues for activation include Thr357/Ser358 in the activation loop and brace helix residues that maintain autoinhibitory interaction between the 4HB and PsKD. Upon necroptotic signaling, RIPK3 phosphorylates MLKL at Thr357/Ser358 within the necrosome complex (RIPK1/RIPK3/MLKL), relieving autoinhibition, promoting MLKL tetramerization or oligomerization, and enabling its translocation to the plasma membrane inner leaflet via PIP2 binding. There, MLKL forms pores that mediate ion influx, plasma membrane rupture, and lytic cell death. MLKL is integral to inflammatory signaling pathways (such as TNF/NF-κB) and distinguishes necroptosis from apoptosis. Dysregulation of MLKL-mediated necroptosis contributes to diseases including inflammatory bowel disease, sepsis, and neurodegeneration (e.g., ALS via neuronal cell death).

Background

MLKL (Mixed Lineage Kinase Domain-Like protein) is a pseudokinase of the protein kinase superfamily that acts as the terminal effector of necroptosis, a form of regulated necrotic cell death triggered by TNF or TLR ligands when caspases are inhibited. MLKL consists of an N-terminal four-helix bundle (4HB) executioner domain (α1–α4 helices that form membrane-disrupting pores), a central two-helix “brace” region connecting domains, and a C-terminal pseudokinase domain (PsKD) that, despite lacking catalytic activity, binds nucleotides and regulates activation. Key residues for activation include Thr357/Ser358 in the activation loop and brace helix residues that maintain autoinhibitory interaction between the 4HB and PsKD. Upon necroptotic signaling, RIPK3 phosphorylates MLKL at Thr357/Ser358 within the necrosome complex (RIPK1/RIPK3/MLKL), relieving autoinhibition, promoting MLKL tetramerization or oligomerization, and enabling its translocation to the plasma membrane inner leaflet via PIP2 binding. There, MLKL forms pores that mediate ion influx, plasma membrane rupture, and lytic cell death. MLKL is integral to inflammatory signaling pathways (such as TNF/NF-κB) and distinguishes necroptosis from apoptosis. Dysregulation of MLKL-mediated necroptosis contributes to diseases including inflammatory bowel disease, sepsis, and neurodegeneration (e.g., ALS via neuronal cell death).

References
[1] Johnston A, et al. J Nat Sci. 2018 Jul;4(7):e513. [2] Grootjans S, et al. Cell Death Differ. 2017 Jul;24(7):1184-1195.

PVDFメンブレン孔径参考表

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%