重组Anti-S6K1抗体[E175] (ab32359)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E175] to S6K1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-S6K1抗体[E175]
参阅全部 S6K1 一抗 -
描述
兔单克隆抗体[E175] to S6K1 -
宿主
Rabbit -
特异性
This antibody may detect both phosphorylated and non-phosphorylated forms of p70 S6 Kinase.
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经测试应用
适用于: WB, IHC-Pmore details
不适用于: Flow Cyt or ICC/IF -
种属反应性
与反应: Mouse, Human
预测可用于: Rat, Cow -
免疫原
Synthetic peptide within Human S6K1. The exact sequence is proprietary.
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阳性对照
- Jurkat cell lysate; human breast carcinoma WB: C-Myc/DDK tagged human S6K1 recombinant protein, C-Myc/DDK tagged human S6K2 recombinant protein.
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常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
存储溶液
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
E175 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32359于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
WB | (1) |
1/1000 - 1/5000. Detects a band of approximately 70 kDa (predicted molecular weight: 59 kDa).
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IHC-P | (2) |
1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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说明 |
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WB
1/1000 - 1/5000. Detects a band of approximately 70 kDa (predicted molecular weight: 59 kDa). |
IHC-P
1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
靶标
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功能
Acts to integrate nutrient and growth factor signals in regulation of protein synthesis, cell proliferation, cell growth, cell cycle progression and cell survival. Downstream effector of the mTOR signaling pathway. Phosphorylates specifically ribosomal protein S6 in response to insulin or several classes of mitogens. During translation initiation, the inactive form associatess with the eIF-3 complex under conditions of nutrient depletion. Mitogenic stimulation leads to phosphorylation and dissociation from the eIF-3 complex and the free activated form can phosphorylate other translational targets including EIF4B. Promotes protein synthesis by phosphorylating PDCD4 at 'Ser-67' and targeting it for degradation. Phosphorylates RICTOR leading to regulation of mammalian target of rapamycin complex 2 (mTORC2) signaling; probably phosphorylates RICTOR at 'Thr-1135'. Phosphorylates IRS1 at multiple serine residues coupled with insulin resistance; probably phosphorylates IRS1 at 'Ser-270'. Required for TNF-alpha induced IRS-1 degradation. Phosphorylates EEF2K in response to IGF1 and inhibits EEF2K activity. Phosphorylates BAD at 'Ser-99' in response to IGF1 leading to BAD inactivation and inhibition of BAD-induced apoptosis. Phosphorylates mitochondrial RMP leading to dissociation of a RMP:PPP1CC complex; probably phosphorylates RMP at 'Ser-99'. The free mitochondrial PPP1CC can dephosphorylate RPS6KB1 at Thr-412 which is proposed to be a negative feed back mechanism for the RPS6KB1 antiapoptotic function. Phosphorylates GSK3B at 'Ser-9' under conditions leading to loss of the TSC1-TSC2 complex. Phosphorylates POLDIP3. -
组织特异性
Widely expressed. -
序列相似性
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. S6 kinase subfamily.
Contains 1 AGC-kinase C-terminal domain.
Contains 1 protein kinase domain. -
结构域
The autoinhibitory domain is believed to block phosphorylation within the AGC-kinase C-terminal domain and the activation loop.
The TOS (TOR signaling) motif is essential for activation by mTORC1. -
翻译后修饰
Phosphorylation at Thr-412 is regulated by mTORC1. The phosphorylation at this site is maintained by an agonist-dependent autophosphorylation mechanism. -
细胞定位
Cytoplasm; Nucleus. Cytoplasm and Cell junction > synapse > synaptosome. Mitochondrion outer membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 404181 Cow
- Entrez Gene: 6198 Human
- Entrez Gene: 72508 Mouse
- Entrez Gene: 83840 Rat
- Omim: 608938 Human
- SwissProt: P23443 Human
- SwissProt: Q8BSK8 Mouse
- SwissProt: P67999 Rat
see all -
别名
- 70 kDa ribosomal protein S6 kinase 1 antibody
- KS6B1_HUMAN antibody
- p70 alpha antibody
see all
图片
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All lanes : Anti-S6K1 antibody [E175] (ab32359) at 1/1000 dilution
Lane 1 : C-Myc/DDK tagged human S6K1 recombinant protein 10ng
Lane 2 : C-Myc/DDK tagged human S6K2 recombinant protein 10ng
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 59 kDa
Observed band size: 60,75 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking and dilution buffer and concentration: 5% NFDM/TBST.
ab205606 was also used at 1/1000 dilution to detect C-Myc/DDK tagged human S6K1 and S6K2 recombinant proteins shown on the right panel.
ab32539 showed cross reactivity with S6K2 in recombinant protein testing. S6K1 and S6K2 have different observed band size in endogenous and recombinant protein tests.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: S6K1 knockout HAP1 cell lysate (20 µg)
Lane 3: MCF7 cell lysate (20 µg)
Lane 4: HEK293 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab32359 observed at 68 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32359 was shown to recognize S6K1 when S6K1 knockout samples were used, along with additional cross-reactive bands. Wild-type and S6K1 knockout samples were subjected to SDS-PAGE. ab32359 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging. -
Paraffin-embedded human breast carcinoma
ab32359 at 1/50 dilutionPerform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Anti-S6K1 antibody [E175] (ab32359) + HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Predicted band size: 59 kDa
数据表及文件
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SDS download
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Datasheet download
文献 (61)
ab32359 被引用在 61 文献中.
- Wang Y et al. The synergistic antitumor effect of IL-6 neutralization with NVP-BEZ235 in hepatocellular carcinoma. Cell Death Dis 13:146 (2022). PubMed: 35165269
- Zheng H et al. Dysregulated pseudogene BNIP3P1 inhibited cell proliferation and promoted cell apoptosis in preeclampsia by acting as a competing endogenous RNA for BNIP3. Environ Toxicol 37:971-982 (2022). PubMed: 35187800
- Xu MN et al. Enhanced Activation of mTOR Signaling Pathway Was Found in the Hypertrophic and Nodular Lesions of Port Wine Stains. Clin Cosmet Investig Dermatol 15:643-651 (2022). PubMed: 35444442
- Ouyang H et al. Impaired expression of BCAT1 relates to muscle atrophy of mouse model of sarcopenia. BMC Musculoskelet Disord 23:450 (2022). PubMed: 35562710
- Dong X et al. Effect of serotonin on the cell viability of the bovine mammary alveolar cell-T (MAC-T) cell line. J Anim Sci Technol 64:922-936 (2022). PubMed: 36287778