组蛋白H3（核内参）单克隆抗体

商品属性

产品介绍
英文名称Histone H3 Mouse mAb, Nuclear Loading Control

中文名称组蛋白H3（核内参）单克隆抗体

别    名H3 histone family member E pseudogene; H3 histone family, member A; H3/A; H31_HUMAN; H3F3; H3FA; Hist1h3a; HIST1H3B; HIST1H3C; HIST1H3D; HIST1H3E; HIST1H3F; HIST1H3G; HIST1H3H; HIST1H3I; HIST1H3J; HIST3H3; histone 1, H3a; Histone cluster 1, H3a; Histone H3 3 pseudogene; Histone H3.1; Histone H3/a; Histone H3/b; Histone H3/c; Histone H3/d; Histone H3/f; Histone H3/h; Histone H3/i; Histone H3/j; Histone H3/k; Histone H3/l; H3.1; H3/d; H3C1; H3C10; H3C11; H3C12; H3C2; H3C3; H3C4; H3C7; H3C8; H3FD;  Nuclear Loading Control;

产品类型内参抗体

研究领域细胞生物  免疫学  细胞周期蛋白  细胞类型标志物  

抗体来源Mouse

克隆类型Monoclonal

克 隆 号3G1

交叉反应Human,Mouse,Rat (predicted: Hamster,Bee)

产品应用WB=1:2000-20000,IHC-P=1:1000-2000,IHC-F=1:1000-2000,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15 kDa

检测分子量

细胞定位细胞核

性    状Liquid

浓    度1mg/ml

免 疫 原KLH conjugated synthetic peptide derived from human Histone H3

亚    型IgG

纯化方法affinity purified by Protein G

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Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a replication-dependent histone that is a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is located separately from the other H3 genes that are in the histone gene cluster on chromosome 6p22-p21.3. [provided by RefSeq, Aug 2015]

靶点与功能

组蛋白H3是染色质核心组蛋白家族成员（H2A、H2B、H3、H4），分子量约15kDa，富含赖氨酸和精氨酸，通过N端尾部翻译后修饰（如甲基化、乙酰化） 调控基因表达。作为核内参，其表达水平在不同组织、细胞周期及病理状态下高度稳定，尤其适用于核蛋白提取样本的定量校正（如转录因子、组蛋白修饰酶等）。

实验操作关键要点

WB核蛋白检测优化：

样本制备：使用核蛋白提取试剂盒，确保核浆分离彻底（胞质污染标志物α-tubulin <5%）；

上样量控制：核蛋白建议上样量20-30μg（总蛋白需50-100μg），避免条带过饱和；

抗体稀释：抗体1:3000稀释（5%脱脂奶粉+TBST），抗体1:1000稀释，4℃孵育过夜，二抗1:5000室温孵育1小时。

IF核定位验证：

固定与通透：4%多聚甲醛固定15分钟，0.2% Triton X-100通透10分钟（确保抗体进入细胞核）；

染色方案：一抗1:200稀释，37℃孵育1小时，Alexa Fluor 488二抗1:500避光孵育30分钟，DAPI复染细胞核；

结果判断：荧光信号应完全与DAPI共定位，胞质无明显信号（排除胞质污染）。

ChIP实验注意事项：

超声条件：将染色质打断为200-500bp片段（功率30%，超声30秒×10次，间隔30秒）；

抗体用量：5μg抗体/1×10⁶细胞，4℃旋转孵育过夜，Protein A/G磁珠富集DNA；

阴性对照：设置IgG对照和无抗体对照，确保ChIP信号特异性。

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