重组Anti-Glutathione Peroxidase 4抗体[EPNCIR144] (ab125066)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPNCIR144] to Glutathione Peroxidase 4
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Glutathione Peroxidase 4抗体[EPNCIR144]
参阅全部 Glutathione Peroxidase 4 一抗 -
描述
兔单克隆抗体[EPNCIR144] to Glutathione Peroxidase 4 -
宿主
Rabbit -
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 -
免疫原
Synthetic peptide within Mouse Glutathione Peroxidase 4. The exact sequence is proprietary.
Database link: O70325 -
阳性对照
- WB: HeLa, Jurkat cell lysate; HepG2, A549, 293T whole cell lysate; rat heart lysate, mouse ovary, heart and lung lysate; rat, human and mouse testis tissue lysates. ICC/IF: HEK293 and HeLa cells. IHC-P: Human kidney and stomach tissues. Flow Cyt (intra): HeLa cells.
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常规说明
This antibody was developed as part of a collaboration between Epitomics, the National Cancer Institute's Center for Cancer Research and the lab of Dolph Hatfield. View antibodies from NCI Center for Cancer Research Collaboration.
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).
See other anti-rabbit secondary antibodies that can be used with this antibody.
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
性能
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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. Stable for 12 months at -20°C. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPNCIR144 -
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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KO cell lines
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KO cell lysates
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Positive Controls
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab125066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) |
1/400.
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WB | (8) |
1/1000 - 1/10000. Detects a band of approximately 17 kDa (predicted molecular weight: 22 kDa).
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IHC-P | (2) |
1/50 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF | (2) |
Use at an assay dependent concentration.
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说明 |
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Flow Cyt (Intra)
1/400. |
WB
1/1000 - 1/10000. Detects a band of approximately 17 kDa (predicted molecular weight: 22 kDa). |
IHC-P
1/50 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
靶标
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功能
Protects cells against membrane lipid peroxidation and cell death. Required for normal sperm development and male fertility. Could play a major role in protecting mammals from the toxicity of ingested lipid hydroperoxides. Essential for embryonic development. Protects from radiation and oxidative damage. -
组织特异性
Present primarily in testis. -
序列相似性
Belongs to the glutathione peroxidase family. -
细胞定位
Mitochondrion. Cytoplasm. - Information by UniProt
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数据库链接
- Entrez Gene: 2879 Human
- Entrez Gene: 625249 Mouse
- Entrez Gene: 29328 Rat
- Omim: 138322 Human
- SwissProt: P36969 Human
- SwissProt: O70325 Mouse
- SwissProt: P36970 Rat
- Unigene: 433951 Human
see all -
别名
- Glutathione peroxidase 4 antibody
- GPX 4 antibody
- GPX-4 antibody
see all
图片
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All lanes : Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) at 1/1000 dilution
Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : GPX4 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 22 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?Lanes 1 - 3: Merged signal (red and green). Green - ab125066 observed at 20 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab125066 was shown to react with Glutathione Peroxidase 4 in wild-type HeLa cells in Western blot with loss of signal observed in GPX4 knockout cell line ab262509 (knockout cell lysate ab263935). Wild-type HeLa and GPX4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab125066 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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All lanes : Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) at 1/1000 dilution
Lane 1 : 293T (Human embryonic kidney epithelial cell) whole cell lysate
Lane 2 : A549 (Human lung carcinoma epithelial cell) whole cell lysate
Lane 3 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 4 : Rat heart lysate
Lane 5 : Mouse ovary lysate
Lane 6 : Mouse heart lysate
Lane 7 : Mouse lung 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: 22 kDa
Observed band size: 22 kDa
Exposure time: 20 secondsBlocking/Diluting buffer and concentration 5% NFDM/TBST.
ab181602 was used as GAPDH loading control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Unpurified ab125066, at a 1/100 dilution, staining Glutathione Peroxidase 4 in paraffin-embedded human kidney tissue by immunohistochemistry.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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All lanes : Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066) at 1/1000 dilution
Lane 1 : Rat testis tissue lysate
Lane 2 : Mouse testis tissue lysate
Lane 3 : Human testis tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 22 kDa
Observed band size: 19,22 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Diluting buffer and concentration: 5% NFDM/TBST
This antibody can detect 19kda cytoplasmic form and 22kda mitochondrial isoform.
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Immunocytochemistry/ Immunofluorescence - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)
Immunofluorescence staining of HEK293 cells with purified ab125066 at a working dilution of 1/200, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab125066 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Glutathione Peroxidase 4 (red) with ab125066 at a 1/400 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)Immunohistochemical staining of paraffin embedded human stomach with purified ab125066 at a working dilution of 1/50. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunocytochemistry/ Immunofluorescence - Anti-Glutathione Peroxidase 4 antibody [EPNCIR144] (ab125066)This image is courtesy of an Abreview submitted by Kirk McManus
Unpurified ab125066 staining Glutathione Peroxidase 4 in the HeLa cell line from Human cervical cancer by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methanol. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. ab150081 an Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Nuclear staining was carried out with DAPI.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (343)
ab125066 被引用在 343 文献中.
- Liu Q et al. PUM2 aggravates the neuroinflammation and brain damage induced by ischemia-reperfusion through the SLC7A11-dependent inhibition of ferroptosis via suppressing the SIRT1. Mol Cell Biochem 478:609-620 (2023). PubMed: 35997855
- Barayeu U et al. Hydropersulfides inhibit lipid peroxidation and ferroptosis by scavenging radicals. Nat Chem Biol 19:28-37 (2023). PubMed: 36109647
- Lalonde ME et al. Genome-wide CRISPR screens identify ferroptosis as a novel therapeutic vulnerability in acute lymphoblastic leukemia. Haematologica 108:382-393 (2023). PubMed: 36134452
- Jiang Y et al. Adaptor protein HIP-55-mediated signalosome protects against ferroptosis in myocardial infarction. Cell Death Differ 30:825-838 (2023). PubMed: 36639542
- Li D et al. CST1 inhibits ferroptosis and promotes gastric cancer metastasis by regulating GPX4 protein stability via OTUB1. Oncogene 42:83-98 (2023). PubMed: 36369321