| 来源 | 用途 | 交叉反应性 | 理论分子量 | 实际分子量 |
| Rabbit | ICC, MeDIP | H, M, R | _ | _ |
WB,
Western blot; IP, Immunoprecipitation; IF, Immunofluorescence; IHC, Immunohistochemistry; ICC, Immunocytochemistry; FC, Flow Cytometry; ChIP, Chromatin Immunoprecipitation Assay; ChIP-seq, ChIP-sequencing; MeDIP, Methylated DNA Immunopreciptation.
H, Human; M, Mouse; R, Rat; C, Chicken; Cw, Cow; Dg, Dog; Gp, Guinea pig; Hm, Hamster; Hr, Horse; Mk, Monkey; Pg, Pig; Rb, Rabbit; S, Sheep; Z, Zebrafish; All, all species expected.
配套提供了Western一抗稀释液，可以用于Western检测或其它适当用途时的一抗稀释。
建议抗体使用时的稀释比例如下(实际使用时需根据抗原水平的高低作适当调整)：
| WB | IP | IF | IHC | ICC | FC | ChIP | ChIP-seq |
| - | - | - | 1:50-1:200 | 1:50-1:200 | - | - | 1:1000-1:2000 |
抗体详细信息如下::
| About this Antibody | |
| Name | 5-Methylcytosine (5-mC) Rabbit Polyclonal Antibody |
| Category | Rabbit Polyclonal Antibody (pAb); Primary antibody |
| Isotype | IgG |
| Purification method | Affinity purification |
| Positive samples | - |
| Cellular location | - |
| Customer validation | - |
| About the Immunogen | |
| Immunogen | A synthetic peptide of human 5mC |
| Sequence | - |
| Gene ID | _ |
| Swiss Prot | _ |
| Synonyms | 5-Methylcytosine; 5-mC; N5-Methylcytidine; 5mC; 5meC, 5-meC, 5'-methyl-2'-deoxycytidine, 5MedCyd |
| Category | DNA Methylation; Epigenetics |
| Background | Methylation of DNA at cytosine residues is a heritable, epigenetic modification that is critical for proper regulation of gene expression, genomic imprinting, and mammalian development. 5-methylcytosine is a repressive epigenetic mark established de novo by two enzymes, DNMT3a and DNMT3b, and is maintained by DNMT1. 5-methylcytosine was originally thought to be passively depleted during DNA replication. However, subsequent studies have shown that Ten-Eleven Translocation (TET) proteins TET1, TET2, and TET3 can catalyze the oxidation of methylated cytosine to 5-hydroxymethylcytosine (5-hmC). Additionally, TET proteins can further oxidize 5-hmC to form 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC), both of which are excised by thymine-DNA glycosylase (TDG), effectively linking cytosine oxidation to the base excision repair pathway and supporting active cytosine demethylation. Normally DNA methylation occurs in a bimodal fashion, such that CpG dinucleotides are largely methylated across the genome, except in short stretches of CpG-rich sequences associated with gene promoters, known as CpG-islands, where methylation is virtually absent. Cancer cell genomes often undergo global hypomethylation, while CpG-islands become hypermethylated, causing their associated promoters to become repressed. There is evidence that a number of aberrantly hypermethylated CpG-islands found in carcinomas occur at tumor suppressor genes such as RB1, MLH1, and BRCA1. |
包装清单：
| 产品编号 | 产品名称 | 包装 |
| AF5722 | 5-Methylcytosine (5-mC) Rabbit Polyclonal Antibody | 50μl |
| AZ050 | Western一抗稀释液 | 50ml |
| — | 说明书 | 1份 |
保存条件：
5-Methylcytosine (5-mC) Rabbit Polyclonal Antibody -20℃保存，Western一抗稀释液-20℃或4℃保存，一年有效。Western一抗稀释液优先推荐℃保存，长期不使用可以考虑-20℃保存，但冻融可能会导致出现轻微的浑浊和少量不溶物。
注意事项：
如果本抗体用于Western blot (WB)、免疫荧光(IF)、免疫细胞化学(ICC)等实验，请注意回收使用过的稀释抗体。回收的抗体通常至少可以重复使用5-10次。稀释后的抗体，包括已经使用过的稀释抗体，请4℃保存。
回收后重复使用的抗体，使用方法同新鲜稀释的抗体。如果在重复使用过程中发现抗体出现轻微混浊现象，可以10,000g离心1-3分钟，取上清用于后续检测。如果回收的抗体出现明显的絮状物或长霉长菌等情况，则可以考虑废弃该抗体。
提供的Western一抗稀释液也可以用于免疫荧光(IF)、免疫组化(IHC)、免疫细胞化学(ICC)等适当用途。如果希望获得最佳的检测效果，请考虑使用上述检测专用的一抗稀释液。
本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
为了您的安全和健康，请穿实验服并戴一次性手套操作。
使用说明：
请根据抗体的实际用途选择相应的使用方法。
1. Western检测：
a. 按照推荐的稀释比例用碧云天提供的Western一抗稀释液稀释抗体。
b. 把经过封闭的蛋白膜与稀释好的一抗4℃缓慢摇动过夜或室温缓慢摇动2小时，确保稀释的抗体至少能在摇动的瞬间覆盖蛋白膜。
c. 回收稀释的一抗，4℃保存以备下次继续使用。
d. 按照Western的实验步骤进行后续的洗涤、二抗孵育、洗涤和检测等操作。具体操作可以参考如下网页：
http://www.beyotime.com/western.htm
2. 免疫染色：
可以使用碧云天生产的免疫染色一抗稀释液(P0103)稀释抗体，使用后注意回收稀释好的一抗，具体操作可以参考如下网页：http://www.beyotime.com/support/immunol-staining.htm
3. 其它实验操作请自行参考适当的protocol进行。
