重组Anti-Cyclophilin F抗体[EPR11311-121] (ab231155)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR11311-121] to Cyclophilin F
- 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-Cyclophilin F抗体[EPR11311-121]
参阅全部 Cyclophilin F 一抗 -
描述
兔单克隆抗体[EPR11311-121] to Cyclophilin F -
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: Wild type HAP1 whole cell lysate; HEK-293, HEK-293T, Jurkat, MCF7 and HeLa whole cell lysate; Human liver and heart lysate; Mouse heart lysate; Rat liver and heart lysate. IHC-P: Human pancreas and kidney tissue; Human colonic carcinoma tissue; Mouse colon tissue. ICC/IF: Wildtype HAP1 cells; HeLa cells. Flow Cyt (intra): HeLa and Jurkat 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: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% 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
EPR11311-121 -
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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Immunohistochemistry reagents
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab231155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) |
1/600.
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WB |
1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 22 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Use at 1:4000 (human), 1:500 (mouse) dilution. |
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ICC/IF |
1/100.
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说明 |
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Flow Cyt (Intra)
1/600. |
WB
1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 22 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Use at 1:4000 (human), 1:500 (mouse) dilution. |
ICC/IF
1/100. |
靶标
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功能
PPIases accelerate the folding of proteins. It catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides. -
序列相似性
Belongs to the cyclophilin-type PPIase family.
Contains 1 PPIase cyclophilin-type domain. -
细胞定位
Mitochondrion matrix. - Information by UniProt
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数据库链接
- Entrez Gene: 10105 Human
- Entrez Gene: 105675 Mouse
- Entrez Gene: 282819 Rat
- Omim: 604486 Human
- SwissProt: P30405 Human
- SwissProt: Q99KR7 Mouse
- SwissProt: P29117 Rat
- Unigene: 381072 Human
see all -
形式
This gene encodes a 178 aa mature protein that is found in the mitochondrion and may participate in the permeability transition pore. While technically this protein is Cyclophilin F, literature references commonly refer to this protein as 'cyclophilin D’ or ‘CypD’. A different cytoplasmic protein of 370 aa, represented by Entrez GeneID 5481, is identified as Cyclophilin D. This antibody does not react with this 370 aa cytoplasmic protein. -
别名
- Cyclophilin 3 antibody
- cyclophilin D antibody
- Cyclophilin F antibody
see all
图片
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All lanes : Anti-Cyclophilin F antibody [EPR11311-121] (ab231155) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : PPIF knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 22 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab231155 observed at 23 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab231155 was shown to react with Cyclophilin F in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266077 (knockout cell lysate ab257039) was used. Wild-type HEK-293T and PPIF knockout HEK-293T 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. ab231155 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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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Cyclophilin F with ab231155 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 (green). Confocal image showing Cyclophilin F staining and mitochondrial localization in the HeLa cell line. The nuclear counter stain is DAPI (blue).
Mitochondria are stained with ab33985 Anti-COX IV (mouse mAb) - Mitochondrial Marker followed by ab150120 AlexaFluor®594 Goat anti-Mouse secondary both at 1/1000 dilution (red).
-ve control 1: ab231155 (Rabbit anti-Cyclophilin F) at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab33985 Anti-COX IV (mouse mAb) - Mitochondrial Marker at 1/1000 dilution followed by ab150077 (AlexaFluor®488 Goat anti-Rabbit secondary) at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Cyclophilin F with ab231155 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in mouse colon (PMID: 23991223, PMID: 15344907, PMID: 26258444) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Jurkat (human T cell leukemia T lymphocyte) cell line labeling Cyclophilin F with ab231155 at 1/600 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cellsincubated with secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor� 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-Cyclophilin F antibody [EPR11311-121] (ab231155) at 1/1000 dilution
Lane 1 : Wild type HAP1 whole cell lysate
Lane 2 : Cyclophilin F knockout HAP1 whole cell lysate
Lane 3 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate
Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 22 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?
Exposure time: 59 secondsab231155 was shown to specifically react with Cyclophilin F in wild-type HAP1 cells as signal was lost in Cyclophilin F knockout cells. Wild-type and Cyclophilin F knockout samples were subjected to SDS-PAGE. Ab231155 and ab181602 (Human anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/200000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrumentusing the ECL technique.
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All lanes : Anti-Thrombospondin antibody (ab231115) at 1/1000 dilution
Lane 1 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lane 2 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 3 : HEK-293T (human embryonic kidney epithelial cell), whole cell lysate
Lane 4 : Human liver lysate
Lane 5 : Human heart lysate
Lane 6 : Mouse heart lysate
Lane 7 : Rat liver lysate
Lane 8 : Rat heart lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 22 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Exposure times:
Lanes 1-5: 59 seconds
Lane 6: 37 seconds
Lane 7-8:3 minutes
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Wildtype and Cyclophilin F knockout HAP1 (human chronic myelogenous leukemia near-haploid cell line) cells labeling Cyclophilin F with ab231155 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 (green). Confocal image showing Cyclophilin F staining and mitochondrial co-localization in wildtype HAP1 cells, with loss of the signal in the Cyclophin F-knockout HAP1 cells. The nuclear counter stain is DAPI (blue).
Mitochondria are stained with with ab33985 Anti-COX IV (mouse mAb) - Mitochondrial Marker followed by ab150120 AlexaFluor®594 Goat anti-Mouse secondary both at 1/1000 dilution (red).
-ve control 1: ab231155 (Rabbit anti-Cyclophilin F) at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab33985 Anti-COX IV (mouse mAb) - Mitochondrial Marker at 1/1000 dilution followed by ab150077 (AlexaFluor®488 Goat anti-Rabbit secondary) at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded human colonic carcinoma tissue labeling Cyclophilin F with ab231155 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in tumor cells of human colon carcinoma (PMID: 23991223, PMID: 15344907, PMID: 26258444) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Cyclophilin F with ab231155 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in human kidney (PMID: 23991223, PMID: 15344907, PMID: 26258444) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling Cyclophilin F with ab231155 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in human pancreas (PMID: 23991223, PMID: 15344907, PMID: 26258444) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
-
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cell line labeling Cyclophilin F with ab231155 at 1/600 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor� 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
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
Certificate of Compliance
文献 (1)
ab231155 被引用在 1 文献中.
- Yu CH et al. TDP-43 Triggers Mitochondrial DNA Release via mPTP to Activate cGAS/STING in ALS. Cell 183:636-649.e18 (2020). PubMed: 33031745