Tissue/cell: mouse intestine tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-phospho-CDKN1B(Ser178) Polyclonal Antibody, Unconjugated(bs-5326R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
- ￥1180 - 2800
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- bs-5236R
- 2025年10月24日
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50ul/100ul/200ul
| 规格： | 50ul | 产品价格： | ￥1180.0 |
|---|---|---|---|
| 规格： | 100ul | 产品价格： | ￥1980.0 |
| 规格： | 200ul | 产品价格： | ￥2800.0 |
| 产品编号 | bs-5236R |
| 英文名称 | phospho-CDKN1B (Ser178) Rabbit pAb |
| 中文名称 | 磷酸化P27抗体/周期素依赖激酶抑制剂抗体 |
| 英文别名 | p27Kip1(phospho-Ser178); p-p27Kip1(Ser178); p-p27Kip1(S178); CDN1B_HUMAN; AA408329; AI843786; Cdki1b; CDKN 1B; CDKN 4; CDKN1B; CDKN1B; CDKN4; CDKN4; Cyclin Dependent Kinase Inhibitor 1B; Cyclin Dependent Kinase Inhibitor 1B; Cyclin dependent kinase inhibitor p27; Cyclin dependent kinase inhibitor p27; Cyclin-dependent kinase inhibitor 1B(p27, Kip1); Cyclin-dependent kinase inhibitor 1B; Cyclin-dependent kinase inhibitor p27; Cyclin-dependent kinase inhibitor p27 Kip1; KIP 1; KIP1; MEN1B; MEN4; OTTHUMP00000195098; OTTHUMP00000195099; p27; p27 Kip1; P27-like cyclin-dependent kinase inhibitor; P27KIP1. |
| 产品应用 | WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500 Not yet tested in other applications. |
| 交叉反应 | Human, Mouse, Rat (Dog, Pig, Cow, Horse) |
| 抗体来源 | Rabbit |
| 免疫原 | KLH conjugated Synthesised phosphopeptide derived from human CDKN1B around the phosphorylation site of Ser178 |
| 亚型 | IgG |
| 性状 | Liquid |
| 纯化方法 | affinity purified by Protein A |
| 克隆类型 | Polyclonal |
| 理论分子量 | 22 kDa |
| 浓度 | 1mg/ml |
| 储存液 | 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. |
| 研究领域 | Cancer > Cell cycle > Cell cycle inhibitors > Cip/kip Cell Biology > Cell Cycle > Cell Cycle Inhibitors > Cip / Kip Epigenetics and Nuclear Signaling > Cell cycle > Cell Cycle Inhibitors > Cip/Kip |
| 亚基 | Forms a ternary compex with CCNE1/CDK2/CDKN1B. |
| 亚细胞定位 | Nucleus. Cytoplasm. Endosome. Note=Nuclear and cytoplasmic in quiescent cells. AKT-or RSK-mediated phosphorylation on Thr-198, binds 14-3-3, translocates to the cytoplasm and promotes cell cycle progression. Mitogen-activated UHMK1 phosphorylation on Ser-10 also results in translocation to the cytoplasm and cell cycle progression. Phosphorylation on Ser-10 facilitates nuclear export. Translocates to the nucleus on phosphorylation of Tyr-88 and Tyr-89. Colocalizes at the endosome with SNX6; this leads to lysosomal degradation. |
| 组织特异性 | Expressed in all tissues tested. Highest levels in skeletal muscle, lowest in liver and kidney. |
| 翻译后修饰 | Phosphorylated; phosphorylation occurs on serine, threonine and tyrosine residues. Phosphorylation on Ser-10 is the major site of phosphorylation in resting cells, takes place at the G(0)-G(1) phase and leads to protein stability. Phosphorylation on other sites is greatly enhanced by mitogens, growth factors, cMYC and in certain cancer cell lines. The phosphorylated form found in the cytoplasm is inactivate. Phosphorylation on Thr-198 is required for interaction with 14-3-3 proteins. Phosphorylation on Thr-187, by CDK2 leads to protein ubiquitination and proteasomal degradation. Tyrosine phosphorylation promotes this process. Phosphorylation by PKB/AKT1 can be suppressed by LY294002, an inhibitor of the catalytic subunit of PI3K. Phosphorylation on Tyr-88 and Tyr-89 has no effect on binding CDK2, but is required for binding CDK4. Dephosphorylated on tyrosine residues by G-CSF.
Ubiquitinated; in the cytoplasm by the KPC complex (composed of RNF123/KPC1 and UBAC1/KPC2) and, in the nucleus, by SCF(SKP2). The latter requires prior phosphorylation on Thr-187. Ubiquitinated; by a TRIM21-containing SCF(SKP2)-like complex; leads to its degradation. |
| 相似性 | Belongs to the CDI family. |
| 功能 | Important regulator of cell cycle progression. Involved in G1 arrest. Potent inhibitor of cyclin E- and cyclin A-CDK2 complexes. Forms a complex with cyclin type D-CDK4 complexes and is involved in the assembly, stability, and modulation of CCND1-CDK4 complex activation. Acts either as an inhibitor or an activator of cyclin type D-CDK4 complexes depending on its phosphorylation state and/or stoichometry. |
| 保存条件 | Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. |
| 注意事项 | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. |
| 背景资料 | Cell cycle progression is regulated by cyclins and their cognate Cdks. p27 KIP 1 is a cell cycle regulatory mitotic inhibitor of cdk activity. p27 KIP 1 is a candidate tumor suppressor gene, and has been proposed to function as a possible mediator of TGF beta induced G1 arrest. p27 KIP 1 is up regulated in response to antimitogenic stimuli. The increased protein expression of p27 results in cellular arrest by binding to cyclin/Cdk complexes such as cyclin D1/Cdk4. p27 Kip1 is regulated by phosphorylation on serine 10 (S10) and threonine 187 (T187). Phosphorylation by CDK2 on T187 results in ubiquitylation and degradation of p27 Kip 1; while phosphorylation by hKIS on S10 signals the nuclear export to the cytoplasm. |
| 应用 | 推荐稀释比例 |
| {WB} | {1:500-2000} |
| {IHC-P} | {1:100-500} |
| {IHC-F} | {1:100-500} |
| {IF} | {1:100-500} |
Sample:
Lane 1: NIH/3T3 (Mouse) Cell Lysate at 30 ug
Lane 2: Spleen (Rat) Lysate at 40 ug
Lane 3: Thymus (Rat) Lysate at 40 ug
Lane 4: Hela (Human) Cell Lysate at 30 ug
Lane 5: SH-SY5Y (Human) Cell Lysate at 30 ug
Lane 6: THP-1 (Human) Cell Lysate at 30 ug
Lane 7: HepG2 (Human) Cell Lysate at 30 ug
Lane 8: MCF-7 (Human) Cell Lysate at 30 ug
Primary: Anti-phospho-CDKN1B (Ser178) (bs-5236R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 27 kD
Observed band size: 27 kD
Lane 1: NIH/3T3 (Mouse) Cell Lysate at 30 ug
Lane 2: Spleen (Rat) Lysate at 40 ug
Lane 3: Thymus (Rat) Lysate at 40 ug
Lane 4: Hela (Human) Cell Lysate at 30 ug
Lane 5: SH-SY5Y (Human) Cell Lysate at 30 ug
Lane 6: THP-1 (Human) Cell Lysate at 30 ug
Lane 7: HepG2 (Human) Cell Lysate at 30 ug
Lane 8: MCF-7 (Human) Cell Lysate at 30 ug
Primary: Anti-phospho-CDKN1B (Ser178) (bs-5236R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 27 kD
Observed band size: 27 kD
Paraformaldehyde-fixed, paraffin embedded (mouse skeletal muscle); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-CDKN1B (Ser178)) Polyclonal Antibody, Unconjugated (bs-5236R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat skeletal muscle); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (phospho-CDKN1B (Ser178)) Polyclonal Antibody, Unconjugated (bs-5236R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
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文献和实验相关实验
：使用 Anti-phospho-Akt (Ser473) Rabbit mAb 对石蜡包埋的人乳腺癌组织进行免疫组织化学分析。（图 A）使用免疫组化试剂盒M&R HRP/DAB Detection IHC Kit，抗体 1:100 稀释；(图 B) 采用普通免疫组化试剂盒，抗体 1:25 稀释。 图 6 免疫组化实验检测 Erk1/2 表达 注：使用 Anti-Erk1/2 Mouse mAb与p44/42 MAPK （Erk1/2）Rabbit mAb 对正常小鼠心脏组织进行免疫
at 10, 25, and 50 μM). At 24 h post-treatment, celllysates were obtained; SDS-PAGE and Western blotting were performed as described in Section 3, Methods. AR-A014418dose-dependent decreases of phospho-β-catenin (Ser33/37) and phospho-glycogen synthase
(Ser473)(193H12)Rabbit mAb，1:300,稀释，二抗是驴抗兔，1：2000，我是实验室的新手，刚呆俩月，受 此挫折，一头雾水，请过来人或者有经验的同学帮忙解答怎样解决，非常感谢！ 碧峤 pAkt有两个位点，Ser473和Thr308，其中Ser473相对容易显色。如果显色失败，可能的原因:1 上样量过少，我们一般都是30μg；2 一抗孵育时间过短，我们一般24h~48h；3 一抗是否工作有待排除 米宝
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phospho-CDKN1B (Ser178) Rabbit pAb(bs-5236R)-50ul/100ul/200ul
￥1180 - 2800
