广谱细胞角蛋白PCK单克隆抗体

商品属性

产品介绍
英文名称Pan Cytokeratin Mouse mAb

中文名称广谱细胞角蛋白PCK单克隆抗体

别    名pan-cytokeratin; pan-CK; pan CK; P-CK; CK-PAN; wide spectrum Cytokeratin; Cytokeratins; Cytokeratin Pan; [cytokeratins 10,13,14,15,16,18,19].  

研究领域肿瘤  细胞生物  免疫学  

抗体来源Mouse

克隆类型Monoclonal

克 隆 号4C3

交叉反应Human,Mouse,Rat

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.

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

理论分子量42-64 kDa

检测分子量

细胞定位细胞浆

性    状Liquid

浓    度1mg/ml

免 疫 原KLH conjugated synthetic peptide derived from human Pan Cytokeratin

亚    型IgG2b, Kappa

纯化方法affinity purified by Protein A

缓 冲 液Liquid in PBS containing 50% Glycerol, 0.5% BSA and 0.02% Proclin300.

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Cytokeratins are proteins of keratin-containing intermediate filaments found in the intracytoplasmic cytoskeleton of epithelial tissue. The cytokeratins are encoded by a family encompassing 30 genes. Among them, 20 are epithelial genes and the remaining 10 are specific for trichocytes. In the cytoplasm, the keratin filaments conform a complex network which extends from the surface of the nucleus to the cell membrane. Numerous accessory proteins are involved in the genesis and maintenance of such structure. This association between the plasma membrane and the nuclear surface provides important implications for the organization of the cytoplasm and cellular communication mechanisms. Apart from the relatively static functions provided in terms of supporting the nucleus and providing tensile strength to the cell, the cytokeratin networks undergo rapid phosphate exchanges mediated depolymerization, with important implications in the more dynamic cellular processes such as mitosis and post-mitotic period, cell movement and differentiation. Cytokeratins interact with desmosomes and hemidesmosomes, thus collaborating to cell-cell adhesion and basal cell-underlying connective tissue connection.

靶点与功能

PAP（Prostatic Acid Phosphatase，又称PSAP）是由前列腺上皮细胞溶酶体分泌的糖蛋白，核心功能与临床价值包括：

生理作用：催化精液中磷酸酯水解，维持酸性微环境（pH 5.0-6.0），促进精子获能；

临床标志物：血清PAP水平与前列腺癌骨转移呈正相关（灵敏度82%），与PSA联合检测可将诊断特异性提升15%-20%（尤其对PSA 4-10ng/mL“灰区”患者）；

研究价值：通过IHC定位前列腺癌转移灶（如骨、淋巴结）中的PAP表达，辅助与膀胱癌、直肠癌的鉴别诊断（特异性91%）。

实验操作关键要点

ELISA定量检测（血清样本）：

配对方案：重组兔单抗作为捕获抗体（包被浓度2μg/mL），HRP标记羊抗PAP多抗作为检测抗体，形成双抗夹心体系；

样本处理：血清56℃灭活30分钟（去除补体干扰），1:50稀释后37℃孵育2小时，避免高脂样本（甘油三酯会影响抗原抗体结合）；

灵敏度：最低检测限0.08ng/mL，标准曲线范围0.1-100ng/mL（R²≥0.995），适合低浓度转移灶样本检测。

IHC-P前列腺癌组织染色：

抗原修复：柠檬酸盐缓冲液（pH6.0）微波修复15分钟（95℃），避免EDTA缓冲液（会抑制PAP酶活性）；

阳性定位：正常前列腺组织腺泡上皮细胞胞质呈棕黄色颗粒状阳性，癌组织中表达强度与Gleason评分正相关（Gleason 8-10分患者阳性率93%）；

对照设置：必须包含PAP敲除小鼠前列腺组织（阴性对照）和良性增生组织（弱阳性对照）。

WB验证（LNCaP细胞裂解液）：

样本制备：用含1mM PMSF+10mM NaF（磷酸酶抑制剂） 的RIPA裂解液冰上裂解30分钟，12000g离心取上清；

电泳条件：10% SDS-PAGE凝胶，恒压80V浓缩胶、120V分离胶，转膜采用0.45μm PVDF膜（300mA恒流2小时）；

抗体孵育：一抗1:1000稀释（5% BSA+TBST），4℃孵育过夜，曝光时间45秒（50kDa单体条带为主，100kDa二聚体条带较弱）。

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