重组Anti-eIF4EBP1抗体[Y330] (ab32130)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y330] to eIF4EBP1
- Suitable for: WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-eIF4EBP1抗体[Y330]
参阅全部 eIF4EBP1 一抗 -
描述
兔单克隆抗体[Y330] to eIF4EBP1 -
宿主
Rabbit -
特异性
ab32130 recognises 4E-BP1 (eIF4E-binding protein) also known as PHAS. -
经测试应用
适用于: WB, IHC-P, Flow Cyt (Intra)more details
不适用于: ICC/IF -
种属反应性
与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa, HEK-293 cell lysate, HAP1 and K562 cell lysate, IHC-P: Human prostate carcinoma, Flow Cyt (intra): HeLa cells
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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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
Y330 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32130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 |
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WB |
1/1000. Detects a band of approximately 15-20 kDa (predicted molecular weight: 12 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/1000.
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说明 |
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WB
1/1000. Detects a band of approximately 15-20 kDa (predicted molecular weight: 12 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/1000. |
靶标
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功能
Regulates eIF4E activity by preventing its assembly into the eIF4F complex. Mediates the regulation of protein translation by hormones, growth factors and other stimuli that signal through the MAP kinase and mTORC1 pathways. -
序列相似性
Belongs to the eIF4E-binding protein family. -
翻译后修饰
Phosphorylated on serine and threonine residues in response to insulin, EGF and PDGF. Phosphorylation at Thr-37, Thr-46, Ser-65 and Thr-70 is regulated by mTORC1. Phosphorylated upon DNA damage, probably by ATM or ATR. - Information by UniProt
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数据库链接
- Entrez Gene: 1978 Human
- Omim: 602223 Human
- SwissProt: Q13541 Human
- Unigene: 411641 Human
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别名
- 4E-BP1 antibody
- 4EBP1 antibody
- 4EBP1_HUMAN antibody
see all
图片
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All lanes : Anti-eIF4EBP1 antibody [Y330] (ab32130) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : EIF4EBP1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 12 kDa
Observed band size: 13 kDa why is the actual band size different from the predicted?Lanes 1-2: Merged signal (red and green). Green - ab32130 observed at 13 kDa. Red - loading control ab8245 observed at 37 kDa.
ab32130 Anti-eIF4EBP1 antibody [Y330] was shown to specifically react with eIF4EBP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264784 (knockout cell lysate ab257146) was used. Wild-type and eIF4EBP1 knockout samples were subjected to SDS-PAGE. ab32130 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 Dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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ab32130, staining human prostate carcinoma by immunohistochemistry, Paraffin embedded tissue
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling eIF4EBP1 with purified ab32130 at 1/1000 dilution (1 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Left).
Low expression in mitotic phase was consistent with literature. (PMID: 11553333) -
All lanes : Anti-eIF4EBP1 antibody [Y330] (ab32130) at 1/1000 dilution
Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : eIF4EBP1 knockout HAP1 cell lysate
Lane 3 : Hek293 cell lysate
Lane 4 : K562 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 12 kDaLanes 1 - 4: Merged signal (red and green). Green - ab32130 observed at 20 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32130 was shown to specifically react with eIF4EBP1 when eIF4EBP1 knockout samples were used. Wild-type and eIF4EBP1 knockout samples were subjected to SDS-PAGE. ab32130 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
Anti-eIF4EBP1 antibody [Y330] (ab32130) at 1/1000 dilution + K562 cell lysate
Predicted band size: 12 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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SDS download
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Datasheet download
文献 (9)
ab32130 被引用在 9 文献中.
- Liu L et al. Se improves GPX4 expression and SOD activity to alleviate heat-stress-induced ferroptosis-like death in goat mammary epithelial cells. Anim Cells Syst (Seoul) 25:283-295 (2021). PubMed: 34745435
- Guo Y et al. Spontaneous formation of neutrophil extracellular traps is associated with autophagy. Sci Rep 11:24005 (2021). PubMed: 34907287
- Batool A et al. Eukaryotic Initiation Factor 4E phosphorylation acts a switch for its binding to 4E-BP1 and mRNA cap assembly. Biochem Biophys Res Commun 527:489-495 (2020). PubMed: 32336547
- Hori Y et al. Analysis of mTOR pathway expression in lymphatic malformation and related diseases. Pathol Int 70:323-329 (2020). PubMed: 32067331
- Wu X et al. PI3K/AKT/mTOR pathway inhibitors inhibit the growth of melanoma cells with mTOR H2189Y mutations in vitro. Cancer Biol Ther 19:584-589 (2018). PubMed: 29708815
- Wang W et al. Rasfonin promotes autophagy and apoptosis via upregulation of reactive oxygen species (ROS)/JNK pathway. Mycology 7:64-73 (2016). PubMed: 30123617
- Lu Q et al. Akt inhibition attenuates rasfonin-induced autophagy and apoptosis through the glycolytic pathway in renal cancer cells. Cell Death Dis 6:e2005 (2015). Human . PubMed: 26633711
- Hu Y et al. MiR-99a antitumor activity in human breast cancer cells through targeting of mTOR expression. PLoS One 9:e92099 (2014). WB ; Human . PubMed: 24637915
- Fukuda A et al. Growth-dependent podocyte failure causes glomerulosclerosis. J Am Soc Nephrol 23:1351-63 (2012). PubMed: 22773827