高分子量神经丝蛋白重组兔单抗

商品属性

产品介绍
英文名称NF-H Recombinant Rabbit mAb

中文名称高分子量神经丝蛋白重组兔单抗

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;  

抗体来源Rabbit

克隆类型Recombinant

克 隆 号19F20

交叉反应Human,Mouse,Rat

产品应用IHC-P=1:200-500,IHC-F=1:200-500,IF=1:200-500

not yet tested in other applications.

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

理论分子量112 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 NF-H

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 °C 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

中分子量神经丝蛋白（NF-M，Neurofilament Medium）是神经元中间丝网络的核心亚基（分子量约160kDa），与低分子量（NF-L）和高分子量（NF-H）亚基共同构成神经丝，其核心功能包括：

轴突结构维持：通过与NF-L形成异聚体，调控轴突直径（直径与神经传导速度正相关），在有髓鞘神经元中表达量最高；

病理标志物价值：在肌萎缩侧索硬化症（ALS）、阿尔茨海默病（AD） 等神经退行性疾病中，NF-M异常磷酸化或断裂片段释放到脑脊液/血清中，可作为疾病进展和神经元损伤的敏感指标（ALS患者脑脊液NF-M水平较健康人升高3-5倍）；

神经再生评估：外周神经损伤后，NF-M在再生轴突中的重新表达与神经功能恢复程度正相关。

实验操作关键要点

WB检测（脊髓损伤模型）：

样本处理：SD大鼠脊髓组织用含磷酸酶抑制剂（PhosSTOP） 的RIPA裂解液（含1mM PMSF）冰上裂解30分钟，12000g离心取上清；

电泳条件：7.5% SDS-PAGE凝胶（因分子量较大），恒压80V浓缩胶、120V分离胶，转膜使用0.45μm PVDF膜（300mA恒流2.5小时）；

抗体孵育：重组兔单抗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60秒（160kDa处可见清晰条带，磷酸化修饰后条带迁移率降低）。

IHC-P脑组织染色：

抗原修复：柠檬酸盐缓冲液（pH6.0）微波修复20分钟（石蜡切片），神经元轴突信号显著增强；

阳性定位：大脑皮层锥体细胞轴突束呈棕黄色强阳性，海马齿状回神经元弱阳性，胶质细胞阴性（特异性标志）；

病理样本特征：AD患者脑组织中可见NF-M阳性神经原纤维缠结（与tau蛋白共定位）。

IF轴突再生共定位实验：

与β-III Tubulin抗体共染，NF-M（绿色）标记成熟轴突骨架，β-III Tubulin（红色）标记新生神经元，激光共聚焦可观察损伤后轴突再生的“串珠状”结构。

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