重组Anti-eIF5A抗体[EP526Y] (ab32443)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP526Y] to eIF5A
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Related conjugates and formulations
概述
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产品名称
Anti-eIF5A抗体[EP526Y]
参阅全部 eIF5A 一抗 -
描述
兔单克隆抗体[EP526Y] to eIF5A -
宿主
Rabbit -
特异性
This antibody shows low affinity in recognizing eIF5A2 recombinant protein.
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经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, Human, Recombinant fragment -
免疫原
Synthetic peptide within Human eIF5A aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P63241 -
阳性对照
- Jurkat and HeLa whole cell lysate (ab150035) and Adenocarcinoma of human uterus slides.
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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: 0.01% 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
EP526Y -
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
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32443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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Flow Cyt (Intra) |
1/50 - 1/80.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (1) |
1/5000 - 1/10000. Predicted molecular weight: 18 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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ICC/IF |
1/500.
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说明 |
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Flow Cyt (Intra)
1/50 - 1/80. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/5000 - 1/10000. Predicted molecular weight: 18 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. |
ICC/IF
1/500. |
靶标
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功能
mRNA-binding protein involved in translation elongation. Has an important function at the level of mRNA turnover, probably acting downstream of decapping. Involved in actin dynamics and cell cycle progression, mRNA decay and probably in a pathway involved in stress response and maintenance of cell wall integrity. With syntenin SDCBP, functions as a regulator of p53/TP53 and p53/TP53-dependent apoptosis. Regulates also TNF-alpha-mediated apoptosis. Mediates effects of polyamines on neuronal process extension and survival. May play an important role in brain development and function, and in skeletal muscle stem cell differentiation. Also described as a cellular cofactor of human T-cell leukemia virus type I (HTLV-1) Rex protein and of human immunodeficiency virus type 1 (HIV-1) Rev protein, essential for mRNA export of retroviral transcripts. -
组织特异性
Expressed in umbilical vein endothelial cells and several cancer cell lines (at protein level). -
序列相似性
Belongs to the eIF-5A family. -
翻译后修饰
eIF-5A seems to be the only eukaryotic protein to have an hypusine residue which is a post-translational modification of a lysine by the addition of a butylamino group (from spermidine). -
细胞定位
Cytoplasm. Nucleus. Endoplasmic reticulum membrane. Nucleus > nuclear pore complex. Hypusine modification promotes the nuclear export and cytoplasmic localization and there was a dynamic shift in the localization from predominantly cytoplasmic to primarily nuclear under apoptotic inducing conditions. - Information by UniProt
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数据库链接
- Entrez Gene: 1984 Human
- Entrez Gene: 276770 Mouse
- Entrez Gene: 287444 Rat
- Omim: 600187 Human
- SwissProt: P63241 Human
- SwissProt: P63242 Mouse
- SwissProt: Q3T1J1 Rat
- Unigene: 534314 Human
see all -
别名
- eIF 4D antibody
- eIF 5A 1 antibody
- EIF 5A antibody
see all
图片
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All lanes : Anti-eIF5A antibody [EP526Y] (ab32443) at 1/1000 dilution
Lane 1 : Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate
Lane 2 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 3 : RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 4 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 5 : PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate
Lane 6 : Mouse pancreas tissue lysate
Lane 7 : Mouse heart tissue lysate
Lane 8 : Rat pancreas tissue lysate
Lane 9 : Rat heart tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 18 kDa
Observed band size: 16 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking and Diluting buffer and concentration: 5% NFDM/TBST.
Low expression control: mouse and rat heart.
In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF5A antibody [EP526Y] (ab32443)
Immunohistochemical analysis of paraffin-embedded Rat pancreas labeling eIF5A with ab32443 at 1/1000 dilution (0.101 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on the Rat pancreas.
The section was incubated with ab32443 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF5A antibody [EP526Y] (ab32443)
Immunohistochemical analysis of paraffin-embedded Mouse pancreas labeling eIF5A with ab32443 at 1/1000 dilution (0.101 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on the Mouse pancreas.
The section was incubated with ab32443 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF5A antibody [EP526Y] (ab32443)
Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle labeling eIF5A with ab32443 at 1/1000 dilution (0.101 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: Negligible staining on the mouse cardiac muscle.
The section was incubated with ab32443 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C6 (rat glial tumor glial cell) labelling eIF5A with ab32443 at 1/250 dilution (Green).
Confocal image showing Mitochondria staining in C6 cell line.
Nuclear DNA was labelled with DAPI (shown in blue).
Secondary used was ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 μg/ml).
ab195889 Anti-alpha Tubulin mouse monoclonal antibody -Microtubule Marker (Alexa Fluor® 594) was used as a counterstain at 1/200 (2.5μg/ml) dilution.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) labelling eIF5A with ab32443 at 1/250 dilution (Green).
Confocal image showing Mitochondria staining in NIH/3T3 cell line.
Nuclear DNA was labelled with DAPI (shown in blue).
Secondary used was ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 μg/ml).
ab195889 Anti-alpha Tubulin mouse monoclonal antibody -Microtubule Marker (Alexa Fluor® 594) was used as a counterstain at 1/200 (2.5μg/ml) dilution.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
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Anti-eIF5A antibody [EP526Y] (ab32443) at 1/10000 dilution + Jurkat cell lysate.
Predicted band size: 18 kDa
Observed band size: 18 kDa -
Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling eIF5A with purified ab32443 at 1/500. Cells were fixed with 4% Paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei couterstained with DAPI (blue).
Secondary Only Control: PBS was used instead of the primary antibody as the negative control
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF5A antibody [EP526Y] (ab32443)
ab32443 at a 1:250 dilution staining eIF5A in human adenocarcinoma of uterus using Immunohistochemistry, Paraffin Embedded Tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-eIF5A antibody [EP526Y] (ab32443) at 1/2000 dilution
Lane 1 :Recombinant Human eIF5A protein (ab87457)
Lane 2 :Recombinant Human eIF5A2 protein (ab99140)
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 18 kDa
Observed band size: 15,20 kDa why is the actual band size different from the predicted?
Exposure time: 20 secondsBlocking buffer: 5% NFDM/TBST
This antibody shows low affinity in recognizing eIF5A2 recombinant protein.
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Overlay histogram showing Jurkat cells stained with ab32443 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32443, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (0.5µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
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 (18)
ab32443 被引用在 18 文献中.
- Ding F et al. Circular RNA protein tyrosine kinase 2 aggravates pyroptosis and inflammation in septic lung tissue by promoting microRNA-766/eukaryotic initiation factor 5A axis-mediated ATP efflux. Acta Cir Bras 38:e380323 (2023). PubMed: 36888755
- Choi UY et al. Herpesvirus-induced spermidine synthesis and eIF5A hypusination for viral episomal maintenance. Cell Rep 40:111234 (2022). PubMed: 35977517
- Seoane R et al. eIF5A is activated by virus infection or dsRNA and facilitates virus replication through modulation of interferon production. Front Cell Infect Microbiol 12:960138 (2022). PubMed: 35967877
- Smeltzer S et al. Hypusination of Eif5a regulates cytoplasmic TDP-43 aggregation and accumulation in a stress-induced cellular model. Biochim Biophys Acta Mol Basis Dis 1867:165939 (2021). PubMed: 32882370
- Theodoridis PR et al. Local translation in nuclear condensate amyloid bodies. Proc Natl Acad Sci U S A 118:N/A (2021). PubMed: 33568529