重组Anti-Cytochrome P450 Reductase抗体[EPR14479(B)] (ab180597)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR14479(B)] to Cytochrome P450 Reductase
- 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-Cytochrome P450 Reductase抗体[EPR14479(B)]
参阅全部 Cytochrome P450 Reductase 一抗 -
描述
兔单克隆抗体[EPR14479(B)] to Cytochrome P450 Reductase -
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. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HCT116, HEK-293T, HepG2, HeLa, MCF7 and A431 cell lysates, Mouse and Rat brain, heart, spleen and kidney tissue lysates. ICC/IF: HeLa and MCF7 cells. IHC-P: Human infiltrating duct carcinoma of breast tissue, Mouse and Rat kidney tissue. 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.
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 long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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纯度
Tissue culture supernatant -
克隆
单克隆 -
克隆编号
EPR14479(B) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Assay kits
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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
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab180597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) |
1/10.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/10000 - 1/50000. Detects a band of approximately 77 kDa.
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IHC-P |
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Heat up to 98 °C, below boiling, and then let cool for 10-20 min. |
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ICC/IF |
1/500.
For unpurified use at 1/100. |
说明 |
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Flow Cyt (Intra)
1/10. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/10000 - 1/50000. Detects a band of approximately 77 kDa. |
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Heat up to 98 °C, below boiling, and then let cool for 10-20 min. |
ICC/IF
1/500. For unpurified use at 1/100. |
靶标
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功能
This enzyme is required for electron transfer from NADP to cytochrome P450 in microsomes. It can also provide electron transfer to heme oxygenase and cytochrome B5. -
疾病相关
Defects in POR are the cause of adrenal hyperplasia variant type (AHV) [MIM:201750]; also known as Antley-Bixler syndrome-like phenotype with disordered steroidogenesis. AHV is a rare variant of congenital adrenal hyperplasia. It is an autosomal recessive disorder with apparent combined P450C17 and P450C21 deficiency. Affected girls are born with ambiguous genitalia, but their circulating androgens are low and virilization does not progress. Conversely, affected boys are sometimes born undermasculinized. Boys and girls can also present with bone malformations, in some cases resembling the pattern seen in patients with Antley-Bixler syndrome.
Defects in POR are a cause of isolated disordered steroidogenesis (IDS) [MIM:201750]. -
序列相似性
In the C-terminal section; belongs to the flavoprotein pyridine nucleotide cytochrome reductase family.
Contains 1 FAD-binding FR-type domain.
Contains 1 flavodoxin-like domain. -
细胞定位
Endoplasmic reticulum membrane. Anchored to the ER membrane by its N-terminal hydrophobic region. - Information by UniProt
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数据库链接
- Entrez Gene: 5447 Human
- Entrez Gene: 18984 Mouse
- Entrez Gene: 29441 Rat
- Omim: 124015 Human
- SwissProt: P16435 Human
- SwissProt: P37040 Mouse
- SwissProt: P00388 Rat
- Unigene: 354056 Human
see all -
别名
- CPR antibody
- CYPOR antibody
- Cytochrome p450 oxidoreductase antibody
see all
图片
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Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labelling Cytochrome P450 Reductase with ab180597 at 1/1000 (0.1 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).The section was counterstained with Hematoxylin. 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.
Postive staining on mouse kidney. The section was incubated with ab180597 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labelling Cytochrome P450 Reductase with ab180597 at 1/1000 (0.1 μg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).The section was counterstained with Hematoxylin. 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.
Postive staining on rat kidney. The section was incubated with ab180597 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument -
All lanes : Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] (ab180597) at 1/1000 dilution
Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse heart tissue lysate
Lane 3 : Mouse spleen tissue lysate
Lane 4 : Mouse kidney tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 75 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: Lane 1-3: 26 seconds; Lane 4-7: 75 seconds.
The identities of the lower MW bands between 37 and 60kDa are unknown.
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All lanes : Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] (ab180597) at 1/1000 dilution
Lane 1 : Rat brain tissue lysate
Lane 2 : Rat heart tissue lysate
Lane 3 : Rat spleen tissue lysate
Lane 4 : Rat kidney tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 75 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: Lane 1-3: 26 seconds; Lane 4-7: 75 seconds.
The identities of the lower MW bands between 37 and 60kDa are unknown.
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All lanes : Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] (ab180597) at 1/1000 dilution
Lane 1 : Wild-type HCT116 cell lysate
Lane 2 : POR knockout HCT116 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : A431 cell lysate
Lysates/proteins at 40 µg per lane.
Performed under reducing conditions.
Observed band size: 80 kDa why is the actual band size different from the predicted?Lanes 1- 4: Merged signal (red and green). Green - ab180597 observed at 80 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab180597 was shown to react with Cytochrome P450 Reductase in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266889 (knockout cell lysate ab257596) was used. Wild-type HCT116 and POR knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab180597 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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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Immunohistochemical analysis of paraffin embedded Human infiltrating duct carcinoma of breast tissue labeling Cytochrome P450 Reductase with ab180597 at a 1/150 dilution. HRP Polymer for Rabbit IgG secondary used. Counterstained with Hematoxylin.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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All lanes : Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] (ab180597) at 1/10000 dilution
Lane 1 : Wild-type HeLa lysate
Lane 2 : Cytochrome P450 Reductase knockout HeLa lysate
Lane 3 : HepG2 lysate
Lane 4 : A431 lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.Lanes 1-4: Merged signal (red and green). Green - ab180597 observed at 75 kDa. Red - loading control ab8245 observed at 37 kDa.
ab180597 Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] was shown to specifically react with Cytochrome P450 Reductase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264996 (knockout cell lysate ab257595) was used. Wild-type and Cytochrome P450 Reductase knockout samples were subjected to SDS-PAGE. ab180597 and Anti-GAPDH antibody [6C5] - Loading Control?(ab8245) were incubated overnight at 4^°C at 1 in 10000 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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All lanes : Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] (ab180597) at 1/10000 dilution
Lane 1 : HeLa lysate
Lane 2 : MCF7 lysate
Lane 3 : A431 lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (H+L) Peroxidase conjugated at 1/1000 dilution -
Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling Cytochrome P450 Reductase with purified ab180597 at 1/500 dilution. Cells were fixed with 100% methanol. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.
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Anti-Cytochrome P450 Reductase antibody [EPR14479(B)] (ab180597) at 1/50000 dilution + HepG2 lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG (H+L) Peroxidase conjugated at 1/1000 dilution -
Immunofluorescence analysis of Hela cells (fixative 4% paraformaldehyde) labeling Cytochrome P450 Reductase with ab180597 at a 1/100 dilution (red), and counterstained with Dapi (blue). Goat anti rabbit IgG (Dylight 555) secondary used at a 1/200 diution.
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Immunofluorescence analysis of MCF7 cells (fixative 4% paraformaldehyde) labeling Cytochrome P450 Reductase with ab180597 at a 1/100 dilution (green), and counterstained with Dapi (blue). Goat anti rabbit IgG (Dylight 555) secondary used at a 1/200 diution.
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Intracellular Flow Cytometry analysis of HeLa cells using ab180597 at a 1/10 dilution (red) and a rabbit IgG as negative control (green). Goat anti rabbit IgG (FITC) secondary usedat a 1/150 dilution.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (8)
ab180597 被引用在 8 文献中.
- Lidin E et al. Hippocampal Expression of Cytochrome P450 1B1 in Penetrating Traumatic Brain Injury. Int J Mol Sci 23:N/A (2022). PubMed: 35054909
- Huang S et al. Hepatic TGFβr1 Deficiency Attenuates Lipopolysaccharide/D-Galactosamine-Induced Acute Liver Failure Through Inhibiting GSK3β-Nrf2-Mediated Hepatocyte Apoptosis and Ferroptosis. Cell Mol Gastroenterol Hepatol 13:1649-1672 (2022). PubMed: 35202887
- Wang F et al. PALP: A rapid imaging technique for stratifying ferroptosis sensitivity in normal and tumor tissues in situ. Cell Chem Biol 29:157-170.e6 (2022). PubMed: 34813762
- Bagayoko S et al. Host phospholipid peroxidation fuels ExoU-dependent cell necrosis and supports Pseudomonas aeruginosa-driven pathology. PLoS Pathog 17:e1009927 (2021). PubMed: 34516571
- Yan B et al. Membrane Damage during Ferroptosis Is Caused by Oxidation of Phospholipids Catalyzed by the Oxidoreductases POR and CYB5R1. Mol Cell 81:355-369.e10 (2021). PubMed: 33321093
- Tang M et al. Hippocampal proteomic changes of susceptibility and resilience to depression or anxiety in a rat model of chronic mild stress. Transl Psychiatry 9:260 (2019). PubMed: 31624233
- Storman EM et al. Physical Linkage of Estrogen Receptor a and Aromatase in Rat: Oligocrine and Endocrine Actions of CNS-Produced Estrogens. Endocrinology 159:2683-2697 (2018). PubMed: 29771302
- Reczek CR et al. A CRISPR screen identifies a pathway required for paraquat-induced cell death. Nat Chem Biol 13:1274-1279 (2017). WB ; Human . PubMed: 29058724