高分子量神经丝蛋白抗体

商品属性

产品介绍
英文名称NF-H Rabbit pAb

中文名称高分子量神经丝蛋白抗体

别    名NFH_HUMAN; Neurofilament heavy polypeptide; NEFH; KIAA0845; NFH; NF H; NF200; NF-200; Neurofilament 200; 200 kDa neurofilament protein; Neurofilament triplet H protein; Neurofilament H; Neurofilament heavy polypeptide 200kD;  

研究领域细胞生物  神经生物学  信号转导  细胞凋亡  转录调节因子  

抗体来源Rabbit

克隆类型Polyclonal

克 隆 号

交叉反应Mouse,Rat (predicted: Human)

产品应用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118 kDa

检测分子量200

细胞定位细胞浆

性    状Liquid

浓    度1mg/ml

免 疫 原KLH conjugated synthetic peptide derived from human NF-H: 901-1000/1026

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Neurofilaments can be defined as the intermediate or 10nm filaments found in specifically in neuronal cells. When visualised using an electron microscope, neurofilaments appear as 10nm diameter fibres of indeterminate length that generally have fine wispy protrusions from their sides. They are particularly abundant in axons of large projection neurons. They probably function to provide structural support for neurons and their synapses and to support the large axon diameters required for rapid conduction of impulses down axons. Neurofilaments are composed of a mixture of subunits, which usually includes the three neurofilament triplet proteins neurofilament light (NFL), neurofilament medium (NFM) and neurofilament heavy (NFH). Neurofilaments may also include smaller amounts of peripherin, alpha internexin, nestin and in some cases vimentin. Antibodies to the various neurofilament subunits are very useful cell type markers since the proteins are among the most abundant of the nervous system, are expressed only in neurons, and are biochemically very stable. Some studies have shown that levels of neurofilament heavy and neurofilament light are elevated in patients with Alzheimer's disease, frontotemporal lobe dementia, and vascular dementia.

靶点与功能

NF-κB是一类核转录因子家族，主要包括 p65（RelA）、p50、p52、c-Rel、RelB 等亚基，其中 p65/p50异二聚体 是最经典的活性形式，核心功能包括：

炎症调控：响应TNF-α、LPS等刺激，激活IL-6、TNF-α等促炎因子表达，是关节炎、肠炎等炎症疾病的关键驱动因子；

肿瘤进展：在肝癌（60%）、胃癌（55%） 中持续激活，通过上调抗凋亡基因（Bcl-2）、血管生成因子（VEGF）促进肿瘤增殖与转移；

免疫逃逸：调控PD-L1、CTLA-4等免疫检查点分子表达，帮助肿瘤细胞逃避免疫监视（与PD-1抑制剂耐药相关）。

实验操作关键要点

WB检测p65核转位（炎症模型）：

样本处理：LPS刺激RAW264.7细胞（1μg/mL，6小时），使用核质分离试剂盒分离胞质/核蛋白，核组分中p65条带显著增强；

抗体组合：总p65抗体检测65kDa条带，磷酸化p65抗体检测Ser536磷酸化水平，β-actin（胞质内参）和Lamin B1（核内参）验证分离效果。

IHC-P组织染色（结直肠癌样本）：

抗原修复：柠檬酸盐缓冲液（pH6.0）微波修复20分钟（石蜡切片），肿瘤细胞核信号显著；

阳性定位：正常黏膜上皮细胞胞质弱阳性，癌组织中细胞核呈棕黄色强阳性（与肿瘤分级正相关）；

对照设置：NFKB1 KO小鼠脾脏组织（阴性对照）和LPS刺激小鼠肝脏组织（阳性对照）。

ChIP-qPCR验证DNA结合活性：

抗体用量：5μg抗体/2×10⁶细胞，4℃旋转孵育过夜；

引物设计：针对IL-6启动子区NF-κB结合位点（5'-GGGGAATTTCC-3'），qPCR检测富集倍数（LPS刺激后富集≥20倍）。

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