RIG-I-MAVS 信号通路-免疫学专题- ABclonal

RIG-I-MAVS 信号通路

作为首要的胞浆RNA监测机制，RIG-I-MAVS信号通路被作为一种抗病毒感染的天然免疫保护系统受到深入研究 [1]。RIG-I样受体（RLRs），包括RIG-I、MDA5和LGP2，是检测病毒感染或复制过程中积累的多种病毒RNA的一系列胞浆RNA解旋酶 [2]。在除pDCs以外的大多数细胞类型中，RLRs对于抗病毒反应是必不可少的。 RIG-I和MDA5表现出不同的配体特异性，并对不同的病毒作出反应，而LGP2促进或拮抗MDA5和RIG-I对病毒RNA的识别 [3]。进一步的研究还发现了其他病毒感受器，如OAS1，它是干扰素诱导下可以被dsRNA激活的抗病毒分子酶，在细胞天然抗病毒反应中起着至关重要的作用。这种蛋白质激活潜在的RNaseL，从而导致病毒RNA降解和抑制病毒复制 [4]。
当RIG-I或MDA5识别其配体时，它与位于线粒体外膜的MAVS蛋白迅速结合。 MAVS作为RLR信号传递的核心接头蛋白，在控制下游信号传递中发挥作用。二聚化的MAVS直接招募TRAF2、TRAF3、TRAF5、TRAF6、CARD9和TRADD等分子，作为在线粒体外膜组装信号复合物的平台[5]。这些信号复合体包含由TRAFs或TRADD进一步招募的其他多种蛋白，包括CIAP1/2、TANK-NAP1-Tbkbp1、TAB2/3-TAB1-TAK1、TBK1-IK Kε、IK Kα-IK Kβ-IK Kγ、和RIP1-FADD-Casp8-Casp10 [6]。通过这些信号复合体，RIG-I-MAVS信号级联最终导致IR F3，IRF7，NF-κB和AP-1的磷酸化和激活，它们共同作用于诱导干扰素和促炎细胞因子的表达。与许多信号通路一样，RIG-I-MAVS信号通路也存在精密的调控方式来实现信号的平衡，包括信号蛋白的翻译后修饰（如泛素化、磷酸化修饰）调节或竞争性蛋白相互作用[7]。此外，RIG-I-MAVS信号通路还会与cGAS-STING[8]、炎症小体[9]和自噬[10，11]等多条信号通路发生串话，增加了RIG-I-MAVS信号调节和功能的复杂性。

ABclonal针对RIG-I-MAVS 信号通路研究，提供以下常见核心靶点的产品

COX5B Rabbit pAb

Western blot analysis of various lysates using COX5B Rabbit pAb (A2640) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates / proteins: 25 μg per lane.
Blocking buffer: 3 % nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

Immunohistochemistry analysis of paraffin-embedded Human colon tissue using COX5B Rabbit pAb (A2640) at a dilution of 1:50 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human liver cancer tissue using COX5B Rabbit pAb (A2640) at a dilution of 1:50 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human liver tissue using COX5B Rabbit pAb (A2640) at a dilution of 1:50 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunofluorescence analysis of NIH/3T3 cells using COX5B Rabbit pAb (A2640) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of PC-12 cells using COX5B Rabbit pAb (A2640) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Rig-I/DDX58 Rabbit pAb

Western blot analysis of extracts of various cell lines, using Rig-I/DDX58 antibody (A13407) 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 Enhanced Kit (RM00021).
Exposure time: 90s.

Immunofluorescence analysis of H9C2 cells using Rig-I/DDX58 antibody (A13407) at dilution of 1:100. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of L929 cells using Rig-I/DDX58 antibody (A13407) at dilution of 1:100. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of U20S cells using Rig-I/DDX58 antibody (A13407) at dilution of 1:100. Blue: DAPI for nuclear staining.

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.

Phospho-NF-kB p65/RelA-S536 Rabbit mAb

Western blot analysis of various lysates, using Phospho-NF-kB p65/RelA-S536 Rabbit mAb (AP1294) at1:1000 dilution (upper) or [KO Validated] NF-kB p65/RelA Rabbit mAb (A22331) at1:50000 dilution (lower) incubated overnight at 4℃. HeLa cells were treated by Calyculin A (100 nM) at 37℃ for 30 minutes after serum-starvation overnight.C6 cells were treated by TNF-α (20 ng/ml) and Calyculin A (50 nM) at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

Western blot analysis of lysates from NIH/3T3 cells using Phospho-NF-kB p65/RelA-S536 Rabbit mAb (AP1294) at 1:1000 dilution incubated overnight at 4℃. NIH/3T3 cells were treated by TNF-α (20 ng/ml) at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3 % nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 180s.

Western blot analysis of lysates from HeLa cells using Phospho-NF-kB p65/RelA-S536 Rabbit mAb (AP1294) at 1:19000 dilution incubated at room temperature for 1.5 hours. HeLa cells were treated by TNF-α (20 ng/ml) and Calyculin A (100 nM) at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3 % nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

Western blot analysis of lysates from C6 cells using Phospho-NF-kB p65/RelA-S536 Rabbit mAb (AP1294) at 1:19000 dilution incubated at room temperature for 1.5 hours. C6 cells were treated by Calyculin A (100 nM) at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3 % nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

SMURF2 Rabbit mAb

Western blot analysis of various lysates using SMURF2 Rabbit mAb (A2278) at 1:1000 dilution.
Secondary antibody: HRP-conjugated 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 Enhanced Kit (RM00021).
Exposure time: 3min.

Immunohistochemistry analysis of paraffin-embedded Human colon using SMURF2 Rabbit mAb (A2278) at dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M Tris/EDTA Buffer (pH 9.0) prior to IHC staining.

Immunofluorescence analysis of NIH-3T3 cells using SMURF2 Rabbit mAb (A2278) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunoprecipitation analysis of 300 μg extracts of Mouse lung using 3 μg SMURF2 antibody (A2278). Western blot was performed from the immunoprecipitate using SMURF2 antibody (A2278) at a dilution of 1:1000.

IRF3 Rabbit pAb

Western blot analysis of lysates from 293T cells using IRF3 Rabbit pAb (A11118) at 1:1000 dilution.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 1s.

Immunohistochemistry analysis of paraffin-embedded Human lung cancer using IRF3 Rabbit pAb (A11118) at dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M PBS Buffer (pH 7.2) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human esophageal cancer using IRF3 Rabbit pAb (A11118) at dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M PBS Buffer (pH 7.2) prior to IHC staining.

Immunofluorescence analysis of HeLa cells using IRF3 Rabbit pAb (A11118) at dilution of 1:100 (40x lens). Secondary antibody: ABflo® 488-conjugated Goat Anti-Rabbit IgG (H+L) (AS073) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunoprecipitation analysis of 300 μg extracts of 293T cells using 3 μg IRF3 antibody (A11118). Western blot was performed from the immunoprecipitate using IRF3 (A11118) at a dilution of 1:1000.

Chromatin immunoprecipitation analysis of extracts of HCT116 cell line, using IRF3 rabbit polyclonal antibody (A11118) and rabbit IgG. The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.

TBK1/NAK Rabbit mAb

Western blot analysis of various lysates using TBK1/NAK Rabbit mAb (A3458) at 1:1000 dilution.
Secondary antibody: HRP-conjugated 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: 3s.

Western blot analysis of various lysates using TBK1/NAK Rabbit mAb (A3458) at 1：1000 dilution.
Secondary antibody: HRP-conjugated 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.

Confocal imaging of MCF7 cells using TBK1/NAK Rabbit mAb (A3458,at dilution of 1:100) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012,dilution 1:400) (Green). DAPI was used for nuclear staining (blue). Objective: 100x.

Confocal imaging of NIH/3T3 cells using TBK1/NAK Rabbit mAb (A3458,at dilution of 1:100) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012,dilution 1:400) (Green). DAPI was used for nuclear staining (blue). Objective: 100x.

Immunoprecipitation analysis of 300 μg extracts from 293T cells using 3 μg TBK1/NAK Rabbit mAb (A3458). Western blot was performed from the immunoprecipitate using TBK1/NAK Rabbit mAb (A3458) at a dilution of 1:1000.

Phospho-TBK1/NAK-S172 Rabbit mAb

Western blot analysis of various lysates using Phospho-TBK1/NAK-S172 Rabbit mAb (AP1026) at 1:1000 dilution. Both HeLa cells and NIH/3T3 cells were treated by Calyculin A (100 nM) at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% BSA.
Detection: ECL Basic Kit (RM00020).
Exposure time: 1min.