产品编号 | 产品名称 | 产品包装 | 产品价格 |
AF1663 | Phospho-PAK1 (Ser144)/PAK2 (Ser141)/PAK3 (Ser139) Rabbit Monoclonal Antibody | 50μl | 709.00元 |
来源 | 用途 | 交叉反应性 | 分子量 |
Rabbit | WB, IP, IF, IHC, ICC, FC | H, M, R | 65.0kDa |
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 | 1:10-1:50 | 1:50-1:200 | 1:50-1:200 | 1:50-1:200 | 1:50-1:100 | - | - |
抗体详细信息如下:
About this Antibody | |
Name | Phospho-PAK1 (Ser144)/PAK2 (Ser141)/PAK3 (Ser139) Rabbit Monoclonal Antibody |
Category | Rabbit Monoclonal Antibody (RabMAb); Primary antibody |
Isotype | IgG |
Purification | Affinity purification |
About the Immunogen | |
Immunogen | Recombinant protein |
Gene ID | 5063/5058/5062(Human); 18481/224105/29431(Mouse); 29433/29432/100910732(Rat) |
SwissProt | O75914/Q13153/Q13177(Human); O88643/Q61036/Q8CIN4(Mouse); P35465/Q62829/Q64303(Rat) |
Synonyms | CDKN1A; MRX30; MRX47; OPHN3; PAK3beta; bPAK; hPAK3 |
Category | Cytoskeletal Signaling |
Background | Three isoforms of serine/threonine kinases, designated aPAK p68, bPAK p65 and gPAK p62, have been shown to exhibit a high degree of sequence homol-ogy with the S. cerevisiae kinase Ste 20, involved in pheromone signaling. The a, b and gPAK isoforms complex specifically with Rac1 and Cdc42 in their active GTP-bound state, inhibiting their intrinsic GTPase activity leading to their autophosphorylation. There are eight sites of autophosphorylation on gPAK, including Ser 19, Ser 141 and Thr 402, and phosphorylation of Ser 141 and Thr 402 is correlated with gPAK activation. Once phosphorylated and their affinity for Rac/Cdc42 reduced, the PAK isoforms disassociate from the complex to seek downstream substrates. One such putative substrate is Mek kinase, an upstream effector of Mek4 which is involved in the JNK signaling pathway. While the PAK isoforms interact in a GTP-dependent manner with Rac1 and Cdc42, they do not interact with Rho. |
包装清单:
产品编号 | 产品名称 | 包装 |
AF1663 | Phospho-PAK1 (Ser144)/PAK2 (Ser141)/PAK3 (Ser139) Rabbit Monoclonal Antibody | 50μl |
AZ050 | Western一抗稀释液 | 50ml |
- | 说明书 | 1份 |
保存条件:
Phospho-PAK1 (Ser144)/PAK2 (Ser141)/PAK3 (Ser139) 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:
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Fig. 1. Western blot analysis of Phospho-PAK1 (Ser144)/PAK2 (Ser141)/PAK3 (Ser139) on different lysates using anti-Phospho-PAK1 (Ser144)/PAK2 (Ser141)/PAK3 (Ser139) antibody at 1/1,000 dilution. Positive control: Lane 1: Hela; Lane 2: NIH/3T3; Lane 3: SH-SY-5Y | Fig. 2. ICC staining Phospho-PAK1 (Ser144)/PAK2 (Ser141)/PAK3 (Ser139) in NIH/3T3 cells (green). The nuclear counter stain is DAPI (blue). Cellswere fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS. |
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Fig. 3. Flow cytometric analysis of NIH/3T3 cells with Phospho-PAK1 (Ser144)/PAK2 (Ser141)/PAK3 (Ser139) 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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