Tissue/cell: rat brain tissue;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Phospho-IRF3(Ser396) Polyclonal Antibody, Unconjugated(bs-3195R) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated (bs-0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,C-0033) was used to stain the cell nuclei
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- bs-3195R
- 2025年10月16日
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| 规格： | 50ul | 产品价格： | ￥1180.0 |
|---|---|---|---|
| 规格： | 100ul | 产品价格： | ￥1980.0 |
| 规格： | 200ul | 产品价格： | ￥2800.0 |
| 产品编号 | bs-3195R |
| 英文名称 | phospho-IRF3 (Ser396) Rabbit pAb |
| 中文名称 | 磷酸化干扰素调节因子3 |
| 英文别名 | IRF3(phospho S396); IRF3(phospho Ser396); p-IRF3(Ser396); p-IRF3(S396); Interferon regulatory factor 3; IRF 3; IRF-3; IRF3; IRF3_HUMAN; MGC94729. |
| 产品应用 | WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500 Not yet tested in other applications. |
| 交叉反应 | Human, Mouse, Rat (Dog, Pig, Cow, Sheep) |
| 抗体来源 | Rabbit |
| 免疫原 | KLH conjugated synthesised phosphopeptide derived from human IRF3 around the phosphorylation site of Ser396 |
| 亚型 | IgG |
| 性状 | Liquid |
| 纯化方法 | affinity purified by Protein A |
| 克隆类型 | Polyclonal |
| 理论分子量 | 47 kDa |
| 浓度 | 1mg/ml |
| 储存液 | 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. |
| 研究领域 | Epigenetics and Nuclear Signaling > Transcription > Other factors Immunology > Innate Immunity > Cytokines > Interferons Immunology > Innate Immunity > TLR Signaling Microbiology > Organism > Virus > RNA Virus > ssRNA positive strand virus > SARS Coronavirus |
| 亚基 | Monomer. Homodimer; phosphorylation-induced. Heterodimer with IRF7. Interacts with CREBBP. May interact with MAVS. Interacts with IKBKE and TBK1. Interacts with TICAM1 and TICAM2. Interacts with rotavirus A NSP1 (via C-terminus); this interaction leads to the proteasome-dependent degradation of IRF3. Interacts with RBCK1. Interacts with TRIM21. Interacts with HERC5. |
| 亚细胞定位 | Cytoplasm. Nucleus. Note=Shuttles between cytoplasmic and nuclear compartments, with export being the prevailing effect. When activated, IRF3 interaction with CREBBP prevents its export to the cytoplasm. |
| 组织特异性 | Expressed constitutively in a variety of tissues. |
| 翻译后修饰 | Constitutively phosphorylated on many serines residues. C-terminal serine/threonine cluster is phosphorylated in response of induction by IKBKE and TBK1. Ser-385 and Ser-386 may be specifically phosphorylated in response to induction. An alternate model propose that the five serine/threonine residues between 396 and 405 are phosphorylated in response to a viral infection. Phosphorylation, and subsequent activation of IRF3 is inhibited by vaccinia virus protein E3.
Ubiquitinated; ubiquitination involves RBCK1 leading to proteasomal degradation. Polyubiquitinated; ubiquitination involves TRIM21 leading to proteasomal degradation. ISGylated by HERC5 resulting in sustained IRF3 activation and in the inhibition of IRF3 ubiquitination by disrupting PIN1 binding. The phosphorylation state of IRF3 does not alter ISGylation. |
| 相似性 | Belongs to the IRF family. Contains 1 IRF tryptophan pentad repeat DNA-binding domain. |
| 功能 | Key transcriptional regulator of type I interferon (IFN)-dependent immune responses and plays a critical role in the innate immune response against DNA and RNA viruses. Regulates the transcription of type I IFN genes (IFN-alpha and IFN-beta) and IFN-stimulated genes (ISG) by binding to an interferon-stimulated response element (ISRE) in their promoters. Acts as a more potent activator of the IFN-beta (IFNB) gene than the IFN-alpha (IFNA) gene and plays a critical role in both the early and late phases of the IFNA/B gene induction. Found in an inactive form in the cytoplasm of uninfected cells and following viral infection, double-stranded RNA (dsRNA), or toll-like receptor (TLR) signaling, becomes phosphorylated by IKBKE and TBK1 kinases. This induces a conformational change, leading to its dimerization and nuclear localization and association with CREB binding protein (CREBBP) to form dsRNA-activated factor 1 (DRAF1), a complex which activates the transcription of the type I IFN and ISG genes. Can activate distinct gene expression programs in macrophages and can induce significant apoptosis in primary macrophages. |
| 保存条件 | Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. |
| 注意事项 | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. |
| 背景资料 | This gene encodes a member of the interferon regulatory transcription factor (IRF) family. The encoded protein is found in an inactive cytoplasmic form that upon serine/threonine phosphorylation forms a complex with CREBBP. This complex translocates to the nucleus and activates the transcription of interferons alpha and beta, as well as other interferon-induced genes. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene. [provided by RefSeq, Nov 2011]. |
| 应用 | 推荐稀释比例 |
| {WB} | {1:500-2000} |
| {IHC-P} | {1:100-500} |
| {IHC-F} | {1:100-500} |
| {IF} | {1:100-500} |
Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Phospho-IRF3(Ser396) Polyclonal Antibody, Unconjugated(bs-3195R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Phospho-IRF3(Ser396) Polyclonal Antibody, Unconjugated(bs-3195R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Sample:
Lane 1: Mouse Heart tissue lysates
Lane 2: Mouse Testis tissue lysates
Lane 3: Rat Heart tissue lysates
Lane 4: Rat Testis tissue lysates
Primary: Anti-Phospho-IRF3 (Ser396) (bs-3195R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 47 kDa
Observed band size: 47 kDa
Lane 1: Mouse Heart tissue lysates
Lane 2: Mouse Testis tissue lysates
Lane 3: Rat Heart tissue lysates
Lane 4: Rat Testis tissue lysates
Primary: Anti-Phospho-IRF3 (Ser396) (bs-3195R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 47 kDa
Observed band size: 47 kDa
Paraformaldehyde-fixed, paraffin embedded (human myocardium); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-IRF3 (Ser396)) Polyclonal Antibody, Unconjugated (bs-3195R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-IRF3 (Ser396)) Polyclonal Antibody, Unconjugated (bs-3195R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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文献和实验该产品被引用文献
[IF={{ 8 }}] {Xiaomei Jiang. et al. A pH-Sensitive Nanoparticle as Reactive Oxygen Species Amplifier to Regulate Tumor Microenvironment and Potentiate Tumor Radiotherapy. INT J NANOMED. 2024 Jan 22} {WB} {Mouse}
[IF={{ 5.9 }}] {Jinglong Xue. et al.Paternal BDE-209 exposure disrupts spermatogenesis in mouse offspring via cGAS-STING pathway activation and mitophagy inhibition.Journal of Environmental Sciences.} {Western blot} {Mouse}
[IF={{ 5.61 }}] {Ando, Makoto, et al. "Poly (I: C) impairs NO donor-induced relaxation by overexposure to NO via the NF-kappa B/iNOS pathway in rat superior mesenteric arteries." Free Radical Biology and Medicine (2017).} {WB} {="Rat"}
[IF={{ 5.2 }}] {Tao Lv. et al. USP35 promotes hepatocellular carcinoma progression by protecting PKM2 from ubiquitination‑mediated degradation. INT J ONCOL. 2023 Oct;63(4):1-15} {WB} {Human}
[IF={{ 4.936 }}] {Ding C et al. β-catenin regulates IRF3-mediated innate immune signalling in colorectal cancer.Cell Prolif. 2018 Oct;51(5):e12464.} {IHC} {Human}
相关实验
：使用 Anti-phospho-Akt (Ser473) Rabbit mAb 对石蜡包埋的人乳腺癌组织进行免疫组织化学分析。（图 A）使用免疫组化试剂盒M&R HRP/DAB Detection IHC Kit，抗体 1:100 稀释；(图 B) 采用普通免疫组化试剂盒，抗体 1:25 稀释。 图 6 免疫组化实验检测 Erk1/2 表达 注：使用 Anti-Erk1/2 Mouse mAb与p44/42 MAPK （Erk1/2）Rabbit mAb 对正常小鼠心脏组织进行免疫
at 10, 25, and 50 μM). At 24 h post-treatment, celllysates were obtained; SDS-PAGE and Western blotting were performed as described in Section 3, Methods. AR-A014418dose-dependent decreases of phospho-β-catenin (Ser33/37) and phospho-glycogen synthase
(Ser473)(193H12)Rabbit mAb，1:300,稀释，二抗是驴抗兔，1：2000，我是实验室的新手，刚呆俩月，受 此挫折，一头雾水，请过来人或者有经验的同学帮忙解答怎样解决，非常感谢！ 碧峤 pAkt有两个位点，Ser473和Thr308，其中Ser473相对容易显色。如果显色失败，可能的原因:1 上样量过少，我们一般都是30μg；2 一抗孵育时间过短，我们一般24h~48h；3 一抗是否工作有待排除 米宝
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phospho-IRF3 (Ser396) Rabbit pAb(bs-3195R)-50ul/100ul/200ul
￥1180 - 2800
