来源 | 用途 | 交叉反应性 | 分子量 |
Rabbit | WB, FC, IHC | H, M, R | 65KDa |
WB, Western blot; IP, Immunoprecipitation; IF, Immunofluorescence; IHC, Immunohistochemistry; ICC, Immunocytochemistry; FC, Flow Cytometry; ELISA, Enzyme-linked Immunosorbent Assay; ChIP, Chromatin Immunoprecipitation Assay.
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 | ELISA | ChIP |
1:1000-1:2000 | - | - | 1:100-1:200 | - | 1:50-1:100 | - | - |
抗体详细信息如下:
About this Antibody | |
Name | NF-κB p65 Rabbit Polyclonal Antibody |
Category | Polyclonal antibody(pAb); Primary antibody |
Isotype | IgG |
Purification | Peptide affinity purified |
About the Immunogen | |
Immunogen | This antibody is produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to N-terminal NF-kB p65. |
Gene ID | 5970 |
SwissProt | Q04206 |
Synonyms | NFKB3; Nuclear factor NF-kappa-B p65 subunit; RELA; TF65; Transcription factor p65; p65 |
Category | Immunology and Inflammation |
Background | NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins. NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression . |
包装清单:
产品编号 | 产品名称 | 包装 |
AF0246 | NF-κB p65 Rabbit Polyclonal Antibody | 50μl |
AZ050 | Western一抗稀释液 | 50ml |
— | 说明书 | 1份 |
保存条件:
NF-κB p65 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4℃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/support/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。
4. 代表性图片:
Fig. 1. Western blot analysis of NF-kB p65 on different cell lysates using anti-NF-kB p65 antibody at 1/1000 dilution.Positive control:Lane1: Hela Lane2:A549 Lane3: PC12 Lane 4: Mouse embryonic Lane5: NIH/3T3 stem cell | Fig. 2. Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti- NF-kB p65 antibody. Counter stained with hematoxylin. |
Fig. 3. Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti- NF-kB p65 antibody. Counter stained with hematoxylin. | Fig. 4. Flow cytometric analysis of Hela cells with NF-kB p65 antibody at 1/50 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red).Goat anti rabbit IgG (FITC) was used as the secondary antibody. |