来源 | 用途 | 交叉反应性 | 理论分子量 | 实际分子量 |
Rabbit | WB | H, M, R | 42/49/53/83/84kDa | 90kDa |
WB, Western blot; IP, Immunoprecipitation; IF, Immunofluorescence; IHC, Immunohistochemistry; ICC, Immunocytochemistry; FC, Flow Cytometry; ChIP, Chromatin Immunoprecipitation Assay; ChIP-seq, ChIP-sequencing.
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 |
1:500-1:2000 | - | - | - | - | - | - | - |
抗体详细信息如下:
About this Antibody | |
Name | PI3 Kinase p85α Rabbit Polyclonal Antibody |
Category | Rabbit Polyclonal Antibody (pAb); Primary antibody |
Isotype | IgG |
Purification method | Affinity purification |
Positive samples | SKOV3, Mouse brain, Rat brain |
Cellular location | - |
Customer validation | - |
About the Immunogen | |
Immunogen | Recombinant fusion protein containing a sequence corresponding to amino acids 240-380 of human PI3 Kinase p85 alpha (NP_852556.2). |
Sequence | EKFKREGNEKEIQRIMHNYDKLKSRISEIIDSRRRLEEDLKKQAAEYREIDKRMNSIKPDLIQLRKTRDQYLMWLTQKGVRQKKLNEWLGNENTEDQYSLVEDDEDLPHHDEKTWNVGSSNRNKAENLLRGKRDGTFLVRE |
Gene ID | 5295 |
Swiss Prot | P27986 |
Synonyms | PIK3R1; AGM7; GRB1; IMD36; p85; p85-ALPHA; phosphoinositide-3-kinase regulatory subunit 1 |
Category | PI3K/Akt Signaling; MAPK/Erk in Growth & Differentiation; SAPK/JNK Signaling; PKC Signaling |
Background | Phosphoinositide 3-kinase (PI3K) catalyzes the production of phosphatidylinositol-3,4,5-triphosphate by phosphorylating phosphatidylinositol (PI), phosphatidylinositol-4-phosphate (PIP), and phosphatidylinositol-4,5-bisphosphate (PIP2). Growth factors and hormones trigger this phosphorylation event, which in turn coordinates cell growth, cell cycle entry, cell migration, and cell survival. PTEN reverses this process, and research studies have shown that the PI3K signaling pathway is constitutively activated in human cancers that have loss of function of PTEN. PI3Ks are composed of a catalytic subunit (p110) and a regulatory subunit. Various isoforms of the catalytic subunit (p110α, p110β, p110γ, and p110δ) have been isolated, and the regulatory subunits are p85α, p85β and p55γ. p85α (PIK3R1) is one of the regulatory subunits that associated with the class IA PI 3-Kinases. Composed of one SH3 and two SH2 domains, p85 alpha functions as an adaptor, coupling catalytic p110 to activated receptor tyrosine kinases. Phosphoinositide 3-kinase p85β subunit (PI3-Kinase p85 beta ), also called regulatory subunit 2 (PIK3-R2) is an 85 kDa, 728 amino acid (aa) regulatory subunit of the PI3K complex. It shares ubiquitous expression and overlapping function with the p85 alpha subunit, but the two are differentially phosphorylated and may have different effects on apoptosis. Within aa 516-642, human p85 beta shares 97%, 96% and 68% aa identity with mouse and rat p85 beta and human p85 alpha, respectively. Full-length human p85 alpha and p85 beta share 59% aa identity. p55γ (Protein of 55 kDa, gamma subunit; also PI3-kinase regulatory subunit gamma and p55PIK) is a 55-57 kDa member of the PI3K p85 family of regulatory subunits. It is widely expressed, and serves as a recruitment subunit for the p110 catalytic subunit of PI3 kinase. p55 gamma /p55PIK should not be confused with the 55 kDa alpha splice variant of the p85a gene (75% amino acid [aa] identity). Human p55 gamma is 461 aa in length. It contains a unique 34 aa N-terminus that binds Rb and prevents cycle progression, followed by a Pro-rich region (aa 35-44) and two SH2 domains that bind the p110 catalytic subunit (aa 65-160 and 358-452). Based on mouse, it is likely that alternate start sites at Met8 and Met32 generate 54 and 50 kDa protein products, respectively. There is also a potential for a start site 18 aa upstream of the standard start site, as well as a splice form that shows a deletion of aa 256-314 that may be accompanied by an additional deletion of aa 36-71. Over aa 251-378, human p55 gamma shares 94% aa identity with mouse p55 gamma. |
包装清单:
产品编号 | 产品名称 | 包装 |
AF7742 | PI3 Kinase p85α Rabbit Polyclonal Antibody | 50μl |
AZ050 | Western一抗稀释液 | 50ml |
— | 说明书 | 1份 |
保存条件:
PI3 Kinase p85α Rabbit Polyclonal Antibody -20ºC保存,Western一抗稀释液-20ºC或4ºC保存,一年有效。Western一抗稀释液优先推荐4ºC保存,长期不使用可以考虑-20ºC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。
4. 代表性图片:
Fig. 1. Western blot analysis of extracts of various cell lines, using PI3 Kinase p85 alpha antibody at 1:1000 dilution.
Secondary antibody: HRP-labeled Goat Anti-Rabbit IgG(H+L) (A0208) at 1:10000 dilution.
Lysates/proteins:25µg per lane.
Blocking buffer: QuickBlock™ Blocking Buffer (P0231)
Detection: BeyoECL Star (P0018A).
Exposure time:1s.