坏死性凋亡经典信号通路-程序性细胞死亡专题- ABclonal

坏死性凋亡经典信号通路

坏死性凋亡是当正常凋亡途径被抑制时发生的一种替代性程序性细胞死亡模式，分别与凋亡和坏死的某些特征相似。坏死性凋亡需要这一途径的关键参与者RIPK3及其底物MLKL[1]。发生坏死性凋亡的细胞经历了与凋亡有明显差别的一些非常独特的形态变化，细胞发生快速肿胀，细胞器同时也发生肿胀，而不是细胞凋亡过程中发生的细胞收缩。此外，发生坏死性凋亡的细胞不会在质膜上出泡，而是经历早期质膜破坏、磷脂酰丝氨酸暴露和细胞器破裂，导致细胞内容物泄漏，从而引发炎症[2]。通过配体依赖性细胞表面死亡受体，如Fas、TNF受体1（TNFR1）、IFN受体（IFNR）、Toll样受体（TLR）和细胞内RNA或DNA感受器，可以启动坏死性凋亡途径，而TNFR1介导的坏死性凋亡过程最具代表性[3]。TNF刺激TNFR1有3种功能结果：依赖于不同调节蛋白的组装，这些不同的途径最终导致NF-κB介导的炎症、Caspase-8介导的凋亡，或在Caspase-8抑制条件下激活坏死性凋亡替代途径[3]。当TNFα与TNFR1受体结合时，该受体被发现触发了含有TRADD、FADD、RIPK1、TRAF2/TRAF5和CIAP1/CIAP2的短期膜信号复合体（称为复合物I）的快速形成[1]。在磷酸化TRAF2存在下，促存活的NF-κB活化通路压制促凋亡的Caspase-8的激活[4]。发生在Ser11上的TRA F2磷酸化促进RIPK1泛素化，促进下游IκB激酶（IKK）复合体的募集和激活[5]。这导致NF-κB激活，并同时阻止RIPK1与FADD和Caspase-8 [6,7]相互作用。在这种情况下，Caspase-8和cFLIPL的异二聚体的形成会抑制细胞凋亡，导致Caspase-8无法活化[8]。在细胞死亡诱导条件下，CYLD[9]的去泛素化诱导TRADD和RIPK1与TNFR1的解离，从而导致复合物II的形成。当FADD和Caspase-8被招募到TRADD和RIPK1中形成复合体IIa，通过寡聚和裂解激活Caspase-8[10]，活化的Caspase-8通过抑制RIPK1和RIPK3[11]的活性，诱导外源性细胞凋亡，使坏死性凋亡不能发生。当Caspase-8被抑制或阻断时，RIPK1和RIPK3相互作用导致自体磷酸化、转磷酸化和“坏死性凋亡复合体”（由RIPK1、RIPK3和MLKL组成）的组装，从而进一步启动下游信号，导致坏死性凋亡[12-14]。虽然RIPK3和RIPK1都是诱导坏死性凋亡所必需的，但当RIPK3在细胞中过度表达时，它本身就能促进坏死性凋亡[15]。某些替代受体/配体相互作用，如双链RNA或LPS刺激TLR3/TLR4、或由DAI感知病毒DNA，导致TRIF-RIPK3复合体[16]或DAI-RIPK3复合体[17]和RIPK1非依赖性复合体形成[18]。RIPK3被激活后，通过磷酸化激活假激酶MLKL，在诱导坏死中发挥关键作用。MLKL的作用方式有两种：（1）作为质膜中钠离子或钙离子通道募集的平台[19]；（2）通过与磷酰肌醇磷酸的氨基末端相互作用促进质膜中的孔道形成[20]。

下载通路

ABclonal提供坏死性凋亡信号研究特色产品

RIPK3 Rabbit pAb

Western blot analysis of various lysates, using RIPK3 Rabbit pAb (A5431) at 1:3000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

Immunohistochemistry analysis of paraffin-embedded Human liver cancer using RIPK3 Rabbit pAb (A5431) at dilution of 1:100 (40x lens).Perform microwave antigen retrieval with 10 mM PBS buffer pH 7.2 before commencing with IHC staining protocol.

Immunohistochemistry analysis of paraffin-embedded Mouse pancreas using RIPK3 Rabbit pAb (A5431) at dilution of 1:100 (40x lens).Perform microwave antigen retrieval with 10 mM PBS buffer pH 7.2 before commencing with IHC staining protocol.

Immunofluorescence analysis of C6 cells using RIPK3 Rabbit pAb (A5431) at dilution of 1:50 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of NIH/3T3 cells using RIPK3 Rabbit pAb (A5431) at dilution of 1:50 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of PC-12 cells using RIPK3 Rabbit pAb (A5431) at dilution of 1:50 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of HT-29 cells using RIPK3 Rabbit pAb (A5431) at dilution of 1:100 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of NIH/3T3 cells using RIPK3 Rabbit pAb (A5431) at dilution of 1:100 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of PC-12 cells using RIPK3 Rabbit pAb (A5431) at dilution of 1:100 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

[KO Validated] Caspase-8 Rabbit mAb

Western blot analysis of various lysates using [KO Validated] Caspase-8 Rabbit mAb (A19549) at 1:1000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 3min.

Western blot analysis of lysates from wild type (WT) and caspase-8 knockout (KO) HeLa cells using [KO Validated] Caspase-8 Rabbit mAb (A19549) at 1:1000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:5000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 5% nonfat dry milk in TBST.
Exposure time: 60s.
The data were kindly provided by Dr. Feng Shao, NIBS

RIPK1/RIP Rabbit mAb

Western blot analysis of various lysates using RIPK1/RIP Rabbit mAb (A19580) at 1:1000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 3min.

Immunoprecipitation analysis of 300 μg extracts from HeLa cells using 3 μg RIPK1/RIP Rabbit mAb (A19580). Western blot was performed from the immunoprecipitate using RIPK1/RIP Rabbit mAb (A19580) at a dilution of 1:500.

FADD Rabbit mAb

Western blot analysis of extracts of various cell lines, using FADD antibody (A19049) at 1:1000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 90s.

TNFAIP3 Rabbit mAb

Western blot analysis of various lysates using TNFAIP3 Rabbit mAb (A19128) at 1:1000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 5s.

Immunohistochemistry analysis of paraffin-embedded human colon carcinoma using TNFAIP3 Rabbit mAb (A19128) at dilution of 1:100 (40x lens).Perform microwave antigen retrieval with 10 mM PBS buffer pH 7.2 before commencing with IHC staining protocol.

MLKL Rabbit mAb

Western blot analysis of various lysates using MLKL Rabbit mAb (A21894) at 1:1000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

Immunohistochemistry analysis of paraffin-embedded human esophageal cancer using MLKL Rabbit mAb (A21894) at dilution of 1:100 (40x lens).Perform microwave antigen retrieval with 10 mM Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

TRAF2 Rabbit pAb

Western blot analysis of various lysates using TRAF2 Rabbit pAb (A0962) at 1:1000 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.

Immunohistochemistry analysis of paraffin-embedded mouse brain using TRAF2 Rabbit pAb (A0962) at dilution of 1:200 (40x lens).Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.

Immunohistochemistry analysis of paraffin-embedded mouse spleen using TRAF2 Rabbit pAb (A0962) at dilution of 1:200 (40x lens).Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.

Immunohistochemistry analysis of paraffin-embedded rat spleen using TRAF2 Rabbit pAb (A0962) at dilution of 1:200 (40x lens).Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.

Immunohistochemistry analysis of paraffin-embedded rat testis using TRAF2 Rabbit pAb (A0962) at dilution of 1:200 (40x lens).Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.

cIAP2 Rabbit pAb

Western blot analysis of various lysates using cIAP2 Rabbit pAb (A0833) at 1:500 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 180s.

Immunohistochemistry analysis of paraffin-embedded rat spleen using cIAP2 Rabbit pAb (A0833) at dilution of 1:20 (40x lens).Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.

Immunofluorescence analysis of A-549 cells using cIAP2 Rabbit pAb (A0833) at dilution of 1:50 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of C6 cells using cIAP2 Rabbit pAb (A0833) at dilution of 1:50 (40x lens). Secondary antibody: Cy3 Goat Anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

坏死性凋亡信号通路精选研究产品

分类

研究靶点	产品货号	产品名称	应用	物种

TNFR1及相关蛋白

TNFRSF1A	A1540	TNFR1/TNFRSF1A Rabbit pAb	ELISA,WB,IHC-P,IF/ICC	Human, Mouse, Rat
Tradd	A1145	TRADD Rabbit pAb	ELISA,WB	Human, Mouse
TRAF2	A0962	TRAF2 Rabbit pAb	ELISA,WB,IHC-P	Human, Mouse, Rat
TRAF2	A19129	[KD Validated] TRAF2 Rabbit mAb	ELISA,WB,IP	Human, Mouse, Rat
CYLD	A3821	CYLD Rabbit pAb	ELISA,WB,IHC-P	Human, Mouse, Rat
TNFAIP3	A19128	TNFAIP3 Rabbit mAb	ELISA,WB,IHC-P	Human
BIRC3	A0833	cIAP2 Rabbit pAb	ELISA,WB,IHC-P,IF/ICC	Human, Mouse, Rat

程序性坏死信号复合体关键组分

RIPK1	A19580	RIPK1/RIP Rabbit mAb	ELISA,WB,IP	Human
RIPK1	A7414	RIPK1/RIP Rabbit pAb	ELISA,WB,IHC-P,IF/ICC,IP	Human, Mouse, Rat
RIPK1	AP1448	Phospho-RIPK1-S166 Rabbit mAb	ELISA,WB	Human
RIPK3	A5431	RIPK3 Rabbit pAb	ELISA,WB,IHC-P,IF/ICC	Human, Mouse, Rat
MLKL	A21894	MLKL Rabbit mAb	ELISA,WB,IHC-P	Human, Mouse, Rat

程序性坏死信号复合体相关组分

FADD	A19049	FADD Rabbit mAb	ELISA,WB	Human
FADD	A18044	[KO Validated] FADD Rabbit pAb	ELISA,WB	Human, Mouse, Rat
CFLAR	A2555	cFLIP Rabbit pAb	ELISA,WB	Human, Mouse, Rat
CASP8	A19549	[KO Validated] Caspase-8 Rabbit mAb	ELISA,WB	Human
CASP8	A0215	Caspase-8 Rabbit pAb	ELISA,WB,IHC-P,IF/ICC	Human, Mouse, Rat

缩略语注释:
IFN, interferon
RIPK, Receptor Interacting Serine/Threonine Kinase
TRADD, TNFRSF1A associated protein with death domain
FADD, Fas-associated protein with death domain
TRAF, TNF receptor-associated factor
cIAP, cellular inhibitors of apoptosis
cFLIPL, cellular FLICE (FADD-like IL-1β-converting enzyme)-inhibitory protein long isoform
CYLD, cylindromatosis (turban tumor syndrome) gene
MLKL, mixed lineage kinase domain-like protein
DAI, DNA-dependent activator of IFN-regulatory factor
TRIF, TIR(Toll/interleukin-1 receptor) domain-containing adaptor protein inducing interferon beta

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