p21蛋白重组兔单抗

商品属性

产品介绍
英文名称p21 Recombinant Rabbit mAb

中文名称p21蛋白重组兔单抗

别    名CDN1A_HUMAN; Cyclin-dependent kinase inhibitor 1; CDKN1A; CAP20; CDKN1; CIP1; MDA6; MDA-6; PIC1; SDI1; WAF1; p21CIP1; CDK-interacting protein 1; Melanoma differentiation-associated protein 6(MDA-6);  

研究领域肿瘤  免疫学  信号转导  转录调节因子  肿瘤细胞生物标志物  

抗体来源Rabbit

克隆类型Recombinant

克 隆 号R5H7

交叉反应Human

产品应用WB=1:500-2000,IHC-P=1:50-200,IHC-F=1:50-200,IF=1:50-200,Flow-Cyt=1:50-100,ICC/IF=1:50-200

not yet tested in other applications.

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

理论分子量18 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 CDKN1A: 50-90

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 potent cyclin-dependent kinase inhibitor. The encoded protein binds to and inhibits the activity of cyclin-cyclin-dependent kinase2 or -cyclin-dependent kinase4 complexes, and thus functions as a regulator of cell cycle progression at G1. The expression of this gene is tightly controlled by the tumor suppressor protein p53, through which this protein mediates the p53-dependent cell cycle G1 phase arrest in response to a variety of stress stimuli. This protein can interact with proliferating cell nuclear antigen, a DNA polymerase accessory factor, and plays a regulatory role in S phase DNA replication and DNA damage repair. This protein was reported to be specifically cleaved by CASP3-like caspases, which thus leads to a dramatic activation of cyclin-dependent kinase2, and may be instrumental in the execution of apoptosis following caspase activation. Mice that lack this gene have the ability to regenerate damaged or missing tissue. Multiple alternatively spliced variants have been found for this gene. [provided by RefSeq, Sep 2015]

靶点与功能

P27（CDKN1B，Cyclin-Dependent Kinase Inhibitor 1B）是Cip/Kip家族抑癌蛋白（198个氨基酸，分子量约27kDa），通过以下机制调控细胞周期：

G1期阻滞：与Cyclin E-CDK2、Cyclin A-CDK2复合物结合，抑制激酶活性，阻止Rb蛋白磷酸化，从而阻止细胞从G1期进入S期；

肿瘤抑制作用：在乳腺癌、胃癌、肺癌等多种肿瘤中低表达或缺失，其表达水平与肿瘤分化程度正相关（P27高表达患者5年生存率提高35%）；

非激酶依赖功能：通过与Skp2、CRM1等蛋白相互作用，参与细胞迁移、凋亡调控（如激活JNK通路促进凋亡）。

实验操作关键要点

WB检测（细胞周期同步化样本）：

样本处理：HCT116细胞经血清饥饿（0.5% FBS）48小时诱导G1期阻滞（P27高表达），或用MG132（10μM）处理24小时抑制P27降解；

电泳条件：12% SDS-PAGE凝胶，恒压80V浓缩胶、120V分离胶，转膜使用0.45μm PVDF膜（200mA恒流1小时）；

抗体孵育：一抗1:1000稀释（5% BSA+TBST），4℃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秒（27kDa处可见清晰条带）。

IHC-P肿瘤组织染色：

抗原修复：柠檬酸盐缓冲液（pH6.0）微波修复15分钟（石蜡切片），P27胞质/核信号显著增强；

阳性定位：正常组织中上皮细胞胞质/核呈阳性（如乳腺小叶上皮），肿瘤组织中低表达或缺失（如三阴乳腺癌P27阳性率<15%）；

预后意义：结直肠癌中P27核阳性患者5年无病生存率提高40%（与核定位率正相关）。

IF双标实验（细胞周期调控）：

与Cyclin E抗体共染，P27（绿色）与Cyclin E（红色）在G1期胞质共定位，S期P27降解后Cyclin E进入细胞核，激光共聚焦可观察二者动态相互作用。

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