来源 | 用途 | 交叉反应性 | 分子量 |
Rabbit | WB, IHC, FC | H, M, R | 28.0kDa |
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 |
1:1,000-1:2,000 | - | - | 1:50-1:200 | - | 1:10-1:50 | - | - |
抗体详细信息如下:
About this Antibody | |
Name | 14-3-3 Rabbit Monoclonal Antibody |
Category | Rabbit Monoclonal Antibody (RabMAb); Primary antibody |
Isotype | IgG |
Purification | Affinity purification |
About the Immunogen | |
Immunogen | Recombinant protein |
Gene ID | 7529/7534(Human); 22631/54401(Mouse); 56011/25578(Rat) |
SwissProt | P31946/P63104(Human); P63101/Q9CQV8(Mouse); P35213/P63102(Rat) |
Synonyms | GW128; HEL-S-1; HS1; KCIP-1; YWHAA/14-3-3-zeta; HEL-S-3; HEL4; KCIP-1; YWHAD |
Category | Cytoskeletal Signaling |
Background | The 14-3-3 family of proteins are important regulatory molecules ubiquitiously expressed in all eukarryotic cells which bind to numerous signaling proteins in various pathways driving critical cellular pathways of apoptosis, differentiation and cell cycle. 14-3-3 theta specifically has also been shown to play an important regulatory role in the TLR2 signaling pathways as a negative regulator of TLR2 ligand Pam3CySk4 induced NF-kB activation. 14-3-3 theta has previously been shown to interact with TLR4 ligand and MyD88 dependent phosphorylated PkC epsilon. 14-3-3 theta in the TLR4 signaling pathway is a positive regulator controlling release of IRF3 indcuded pro-inflammatory cytokines RANTES and IP-10. Currently identified by mass spec as part of the TLR2 signaling complex and taken along with TLR4 data, a 14-3-3 theta antibody can be used to examine the different regulatory functions of 14-3-3 theta for different TLRs through its interaction with common or unique TLR signaling adaptor molecules in addition to MyD88 or PkC epsilon such as TRAM or TRIF allowing further clarification of TLR specific pathway regulation. |
包装清单:
产品编号 | 产品名称 | 包装 |
AF2779 | 14-3-3 Rabbit Monoclonal Antibody | 50μl |
AZ050 | Western一抗稀释液 | 50ml |
- | 说明书 | 1份 |
保存条件:
14-3-3 Rabbit Monoclonal 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 14-3-3 on different lysates using anti-14-3-3 antibody at 1/1,000 dilution. Positive control: Lane 1: HT29; Lane 2: Human skin; Lane 3: NIH/3T3 | Fig. 2. Immunohistochemical analysis of paraffin-embedded mouse skin tissue using anti-14-3-3 antibody. Counter stained with hematoxylin. | |
Fig. 3. Flow cytometric analysis of SH-SY-5Y cells with 14-3-3 antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody. |
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