抑癌基因p16重组兔单抗

商品属性

产品介绍
英文名称CDKN2A/p16-INK4a Recombinant Rabbit mAb

中文名称抑癌基因p16重组兔单抗

别    名CDKN2; MTS1; isoforms 1/2/3; Cyclin-dependent kinase 4 inhibitor A; CDK4I; P16INK4; p16INK4a; Multiple tumor suppressor 1;  

抗体来源Rabbit

克隆类型Recombinant

克 隆 号14E19

交叉反应Human

产品应用WB=1:500-2000,IHC-P=1:100-500,IHC-F=1:100-500,IF=1:100-500,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17 kDa

检测分子量

细胞定位细胞核 细胞浆

性    状Liquid

免 疫 原A synthesized peptide derived from human CDKN2A: 120-156/156

亚    型IgG

纯化方法affinity purified by Protein A

缓 冲 液10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% 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 generates several transcript variants which differ in their first exons. At least three alternatively spliced variants encoding distinct proteins have been reported, two of which encode structurally related isoforms known to function as inhibitors of CDK4 kinase. The remaining transcript includes an alternate first exon located 20 Kb upstream of the remainder of the gene; this transcript contains an alternate open reading frame (ARF) that specifies a protein which is structurally unrelated to the products of the other variants. This ARF product functions as a stabilizer of the tumor suppressor protein p53 as it can interact with, and sequester, the E3 ubiquitin-protein ligase MDM2, a protein responsible for the degradation of p53. In spite of the structural and functional differences, the CDK inhibitor isoforms and the ARF product encoded by this gene, through the regulatory roles of CDK4 and p53 in cell cycle G1 progression, share a common functionality in cell cycle G1 control. This gene is frequently mutated or deleted in a wide variety of tumors, and is known to be an important tumor suppressor gene. [provided by RefSeq, Sep 2012]

靶点与功能

p16（CDKN2A/INK4a）是细胞周期依赖性激酶抑制剂，像“分子哨兵”一样通过抑制CDK4/CDK6活性，阻止细胞从G1期进入S期，从而抑制异常增殖。它的核心研究价值在于：

肿瘤诊断：在宫颈癌、黑色素瘤、肺癌等多种肿瘤中因基因甲基化或缺失导致表达下调，是癌前病变筛查的重要标志物（如宫颈上皮内瘤变p16阳性提示进展风险）；

机制研究：通过检测p16表达变化，揭示细胞周期紊乱与肿瘤发生的关联（如p16缺失会解除对Rb通路的抑制，导致细胞无限增殖）。

实验应用关键操作

IHC病理诊断（宫颈癌组织切片）：

抗原修复：柠檬酸钠缓冲液（pH6.0）高温高压修复2分钟，自然冷却后3% H₂O₂阻断内源性过氧化物酶；

阳性定位：正常鳞状上皮胞核阳性，癌前病变/肿瘤组织中因基因沉默呈阴性或弱阳性，可与Ki-67联合染色（p16⁺Ki-67⁺提示高危病变）；

对照设置：用正常皮肤组织作阳性对照（基底细胞胞核强阳性），p16敲除细胞系作阴性对照。

WB验证（A549肺癌细胞系）：

样本处理：RIPA裂解液（含1mM PMSF）冰上裂解30分钟，12000g离心取上清，上样量30μg总蛋白；

关键技巧：使用15% SDS-PAGE凝胶（分离小分子量蛋白），一抗1:1000稀释（5% BSA+TBST），4℃孵育过夜，曝光10-15秒可见16kDa条带。

IF细胞定位（Hela细胞）：

染色步骤：细胞爬片用4%多聚甲醛固定15分钟，0.3% Triton X-100通透后，一抗1:200稀释4℃孵育过夜，Alexa Fluor 488二抗室温孵育1小时；

结果观察：p16在细胞核内呈绿色荧光，与DAPI（蓝色）共定位。

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