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
製品の概要
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製品名
Anti-Glutathione Peroxidase 4 antibody [EPNCIR144]
Glutathione Peroxidase 4 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPNCIR144] to Glutathione Peroxidase 4 -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide within Mouse Glutathione Peroxidase 4. The exact sequence is proprietary.
Database link: O70325 -
ポジティブ・コントロール
- 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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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
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. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPNCIR144 -
アイソタイプ
IgG -
研究分野
関連製品
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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
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab125066の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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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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特記事項 |
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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. |
ターゲット情報
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機能
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. -
組織特異性
Present primarily in testis. -
配列類似性
Belongs to the glutathione peroxidase family. -
細胞内局在
Mitochondrion. Cytoplasm. - Information by UniProt
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参照データベース
- 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 -
別名
- Glutathione peroxidase 4 antibody
- GPX 4 antibody
- GPX-4 antibody
see all
画像
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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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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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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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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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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.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (344)
ab125066 は 344 報の論文で使用されています。
- 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