GADD153重组兔单抗

商品属性

产品介绍
英文名称DDIT3 Recombinant Rabbit mAb

中文名称GADD153重组兔单抗

别    名GADD153; CHOP; Growth arrest and DNA damage-inducible 153; C/EBP homologous protein; C/EBP Homology Protein; CEBPZ; CHOP10; DDIT 3; DNA Damage Inducible Transcript 3; GADD 153; Growth Arrest and DNA Damage Inducible Protein 153; Growth arrest and DNA damage inducible protein GADD153; MGC4154; DDIT3_HUMAN.  

研究领域免疫学  信号转导  细胞凋亡  细胞周期蛋白  转录调节因子  

抗体来源Rabbit

克隆类型Recombinant

克 隆 号6C8

交叉反应Human,Mouse,Rat

产品应用WB=1:1000-2000,IHC-P=1:100-500,IHC-F=1:100-500,IF=1:100-500

not yet tested in other applications.

optimal dilutions/concentrations should be determined by the end user.

理论分子量19 kDa

检测分子量

细胞定位细胞核 细胞浆

性    状Liquid

浓    度1mg/ml

免 疫 原A synthesized peptide derived from human DDIT3: 140-169

亚    型IgG

纯化方法affinity purified by Protein A

缓 冲 液0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

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.

PubMedPubMed

产品介绍This gene encodes a member of the CCAAT/enhancer-binding protein (C/EBP) family of transcription factors. The protein functions as a dominant-negative inhibitor by forming heterodimers with other C/EBP members, such as C/EBP and LAP (liver activator protein), and preventing their DNA binding activity. The protein is implicated in adipogenesis and erythropoiesis, is activated by endoplasmic reticulum stress, and promotes apoptosis. Fusion of this gene and FUS on chromosome 16 or EWSR1 on chromosome 22 induced by translocation generates chimeric proteins in myxoid liposarcomas or Ewing sarcoma. Multiple alternatively spliced transcript variants encoding two isoforms with different length have been identified. [provided by RefSeq, Aug 2010]. Function : Multifunctional transcription factor in ER stress response. Plays an essential role in the response to a wide variety of cell stresses and induces cell cycle arrest and apoptosis in response to ER stress. Plays a dual role both as an inhibitor of CCAAT/enhancer-binding protein (C/EBP) function and as an activator of other genes. Acts as a dominant-negative regulator of C/EBP-induced transcription: dimerizes with members of the C/EBP family, impairs their association with C/EBP binding sites in the promoter regions, and inhibits the expression of C/EBP regulated genes. Positively regulates the transcription of TRIB3, IL6, IL8, IL23, TNFRSF10B/DR5, PPP1R15A/GADD34, BBC3/PUMA, BCL2L11/BIM and ERO1L. Negatively regulates; expression of BCL2 and MYOD1, ATF4-dependent transcriptional activation of asparagine synthetase (ASNS), CEBPA-dependent transcriptional activation of hepcidin (HAMP) and CEBPB-mediated expression of peroxisome proliferator-activated receptor gamma (PPARG). Inhibits the canonical Wnt signaling pathway by binding to TCF7L2/TCF4, impairing its DNA-binding properties and repressing its transcriptional activity. Plays a regulatory role in the inflammatory response through the induction of caspase-11 (CASP4/CASP11) which induces the activation of caspase-1 (CASP1) and both these caspases increase the activation of pro-IL1B to mature IL1B which is involved in the inflammatory response.

靶点与功能

GADD153（Growth Arrest and DNA Damage-inducible Protein 153），又称DDIT3（DNA Damage Inducible Transcript 3），是C/EBP家族转录因子，分子量约32kDa，主要通过以下机制参与细胞应激反应：

内质网应激通路：在未折叠蛋白反应（UPR）中被PERK/eIF2α通路激活，通过抑制抗凋亡基因（如Bcl-2）、激活促凋亡基因（如CHOP）诱导细胞凋亡；

代谢调控：抑制脂质合成关键酶（如SREBP1c），参与非酒精性脂肪肝（NAFLD）的病理过程；

肿瘤微环境：在缺氧、营养剥夺条件下高表达，与乳腺癌、肝癌的化疗耐药性密切相关。

实验操作关键要点

WB检测（内质网应激模型细胞）：

样本处理：用含1mM PMSF的RIPA裂解液（含1% SDS）冰上裂解20分钟，12000g离心15分钟取上清；推荐使用衣霉素（Tunicamycin，5μg/mL）处理HepG2细胞24小时作为阳性对照；

电泳条件：10% SDS-PAGE凝胶，恒压80V浓缩胶、120V分离胶，转膜选用0.45μm PVDF膜（250mA恒流1小时）；

抗体孵育：重组兔单抗推荐1:2000稀释（5% BSA+TBST），4℃摇床孵育过夜，HRP二抗（山羊抗兔）1:5000室温孵育1小时，曝光时间30-60秒（避免与非特异性条带混淆）。

IHC-P组织染色（肝纤维化样本）：

抗原修复：采用柠檬酸钠缓冲液（pH6.0） 微波修复15分钟（95℃），冷却后用3% H₂O₂阻断内源性过氧化物酶10分钟；

阳性定位：肝细胞胞核与胞质均有表达（应激早期胞质为主，晚期核转位），纤维化区域炎症细胞呈强阳性；

封闭处理：5%驴血清+TBST室温封闭30分钟，一抗1:300稀释后4℃孵育过夜，DAB显色控制在3-5分钟。

IF双标实验（内质网共定位）：

推荐与Calnexin抗体（内质网标志物） 共染，使用Alexa Fluor 488（绿色，GADD153）和594（红色，Calnexin），激光共聚焦观察应激条件下二者共定位增强（提示GADD153内质网应激响应）。

公司正在出售的产品