Anti-p27 KIP 1 (phospho T157)抗体(ab85047)
- 数据表
- 具体的参考文献 (1)
- 实验方案
概述
- 产品名称Anti-p27 KIP 1 (phospho T157)抗体
参阅全部 p27 KIP 1 一抗 - 描述兔多克隆抗体to p27 KIP 1 (phospho T157)
- 经测试应用适用于: IHC-P, ICC/IF, WBmore details
- 种属反应性与反应: Human
预测可用于: Cat, Dog, Pig
- 免疫原
Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human p27 KIP 1.
(Peptide available as ab97604.)
- 阳性对照
- This antibody gave a positive signal in the following whole cell lysates: HeLa; Jurkat; Caco2; MCF7.
性能
- 形式Liquid
- 存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
- 存储溶液Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... - 纯度Immunogen affinity purified
- 克隆多克隆
- 同种型IgG
- 研究领域
相关产品
- Compatible Secondaries
应用
Our Abpromise guarantee covers the use of ab85047 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| 应用 | Ab评论 | 说明 |
|---|---|---|
| IHC-P | Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
| ICC/IF | Use a concentration of 5 µg/ml. | |
| WB | Use a concentration of 1 µg/ml. Detects a band of approximately 32 kDa (predicted molecular weight: 22 kDa). |
靶标
- 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.
- 组织特异性Expressed in all tissues tested. Highest levels in skeletal muscle, lowest in liver and kidney.
- 疾病相关Defects in CDKN1B are the cause of multiple endocrine neoplasia type 4 (MEN4) [MIM:610755]. Multiple endocrine neoplasia (MEN) syndromes are inherited cancer syndromes of the thyroid. MEN4 is a MEN-like syndrome with a phenotypic overlap of both MEN1 and MEN2.
- 序列相似性Belongs to the CDI family.
- 结构域A peptide sequence containing only AA 28-79 retains substantial Kip1 cyclin A/CDK2 inhibitory activity.
- 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.
Subject to degradation in the lysosome. Interaction with SNX6 promotes lysosomal degradation. - 细胞定位Nucleus. Cytoplasm. Endosome. 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 and this leads to lysosomal degradation.
- Information by UniProt
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数据库链接
- Entrez Gene: 493958 Cat
- Entrez Gene: 403429 Dog
- Entrez Gene: 1027 Human
- Omim: 600778 Human
- SwissProt: O19001 Cat
- SwissProt: Q6SLL5 Dog
- SwissProt: P46527 Human
- Unigene: 238990 Human
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别名
- AA408329 antibody
- AI843786 antibody
- Cdki1b antibody
see all
Anti-p27 KIP 1 (phospho T157) antibody 图像
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Western blot - p27 KIP 1 (phospho T157) antibody (ab85047)All lanes : Anti-p27 KIP 1 (phospho T157) antibody (ab85047) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg
Lane 3 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate at 10 µg
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg
Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human p27 KIP 1 (phospho T157) peptide (ab97604) at 1 µg/ml
Lane 6 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Human p27 KIP 1 (phospho T157) peptide (ab97604) at 1 µg/ml
Lane 7 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate with Human p27 KIP 1 (phospho T157) peptide (ab97604) at 1 µg/ml
Lane 8 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate with Human p27 KIP 1 (phospho T157) peptide (ab97604) at 1 µg/ml
Lane 9 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with p27 KIP 1 peptide (ab116623) at 1 µg/ml
Lane 10 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with p27 KIP 1 peptide (ab116623) at 1 µg/ml
Lane 11 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate with p27 KIP 1 peptide (ab116623) at 1 µg/ml
Lane 12 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate with p27 KIP 1 peptide (ab116623) at 1 µg/ml
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 22 kDa
Observed band size : 32 kDa (why is the actual band size different from the predicted?)
Additional bands at : 100 kDa,75 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 2 minutes -
Immunocytochemistry/ Immunofluorescence - p27 KIP 1 (phospho T157) antibody (ab85047)ICC/IF image of ab85047 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab85047, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 5µg/ml. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - p27 KIP 1 (phospho T157) antibody (ab85047)IHC image of p27 KIP 1 (phospho T157) staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab85047, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX
Anti-p27 KIP 1 (phospho T157) antibody (ab85047)参考文献
This product has been referenced in:
- Inge LJ et al. Dasatinib, a small molecule inhibitor of the Src kinase, reduces the growth and activates apoptosis in pre-neoplastic Barrett's esophagus cell lines: evidence for a noninvasive treatment of high-grade dysplasia. J Thorac Cardiovasc Surg 145:531-8 (2013). Human . Read more (PubMed: 23142123) »
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