Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
Key features and details
- Mouse monoclonal [E11AE12BD4] to Cyclophilin F
- Suitable for: WB, Flow Cyt, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Cow, Human
- Isotype: IgG1
Related conjugates and formulations
Overview
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Product name
Anti-Cyclophilin F antibody [E11AE12BD4]
See all Cyclophilin F primary antibodies -
Description
Mouse monoclonal [E11AE12BD4] to Cyclophilin F -
Host species
Mouse -
Tested applications
Suitable for: WB, Flow Cyt, ICC/IF, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Cow, Human -
Immunogen
Recombinant full length protein corresponding to Rat Cyclophilin F aa 1-206. (also known as CypD)
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Positive control
- Isolated mitochondria from Human heart, Bovine heart, Rat heart, Mouse heart, HEK293T, HepG2 cells; Cultured Human embryonic lung-derived fibroblasts (strain MRC5); Human cerebellum tissue; HL60 cells.
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General notes
This monoclonal antibody to cyclophilin F has been knockout validated in Western blot. The expected band for cyclophilin F was observed in wild type cells and the band was not seen in knockout cells.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituent: HEPES buffered saline -
Concentration information loading...
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Purification notes
ab110324 was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation. -
Clonality
Monoclonal -
Clone number
E11AE12BD4 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation 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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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab110324 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (2) |
Use a concentration of 1 µg/ml. Predicted molecular weight: 22 kDa.
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Flow Cyt |
Use a concentration of 1 µg/ml.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
Use a concentration of 5 µg/ml.
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IHC-P |
1/100. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
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Notes |
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WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 22 kDa. |
Flow Cyt
Use a concentration of 1 µg/ml. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use a concentration of 5 µg/ml. |
IHC-P
1/100. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. |
Target
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Function
PPIases accelerate the folding of proteins. It catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides. -
Sequence similarities
Belongs to the cyclophilin-type PPIase family.
Contains 1 PPIase cyclophilin-type domain. -
Cellular localization
Mitochondrion matrix. - Information by UniProt
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Database links
- Entrez Gene: 414346 Cow
- Entrez Gene: 10105 Human
- Entrez Gene: 105675 Mouse
- Entrez Gene: 282819 Rat
- Omim: 604486 Human
- SwissProt: P30404 Cow
- SwissProt: P30405 Human
- SwissProt: Q99KR7 Mouse
see all -
Form
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. -
Alternative names
- Cyclophilin 3 antibody
- cyclophilin D antibody
- Cyclophilin F antibody
see all
Images
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Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: Cyclophilin F knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: Hek293 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab110324 observed at 24 kDa. Red - loading control, ab181602, observed at 37 kDa.ab110324 detected the expected band for Cyclophilin F in wild type HAP1 cells and the band was not seen in Cyclophilin F knockout HAP1 cells. Wild-type and Cyclophilin F knockout samples were subjected to SDS-PAGE. Ab110324 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry analysis using ab110324 at 1µg/ml staining Cyclophilin 40 in Cultured Human embryonic lung-derived fibroblasts (strain MRC5), (fixed, treated for heat-induced antigen retrieval, permeabilized) followed by an AlexaFluor® 488-conjugated-goat-anti-mouse IgG1 isotype specific secondary antibody (2 µg/ml).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)Immunohistological analysis using ab110324 at 1µg/ml staining Cyclophilin F in Human cerebellum tissue (Formalin-fixed, Paraffin-embedded).
Note: immunoactivity is most intense in neuronal cell bodies, most notably in the large Purkinje cells. -
All lanes : Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324) at 1/10000 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 - ab110324 observed at 23 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.
ab110324 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. ab110324 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)This image is courtesy of an anonymous AbreviewAll lanes : Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324) at 1/1000 dilution
Lane 1 : WT mouse liver mitochondria lysate at 25 µg
Lane 2 : CypD KO mouse liver mitochondria lysate at 25 µg
Lane 3 : WT mouse liver mitochondria lysate at 35 µg
Lane 4 : CypD KO mouse liver mitochondria lysate at 35 µg
Lane 5 : WT mouse liver mitochondria lysate at 50 µg
Lane 6 : CypD KO mouse liver mitochondria lysate at 50 µg
Secondary
All lanes : HRP-conjugated goat anti-mouse IgG polyclonal at 1/4000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 22 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?
Additional bands at: 13 kDa (possible non-specific binding)
Exposure time: 5 secondsBlocked with 5% milk for 1 hour at 25°C.
Incubated with the primary antibody diluted in PBS-T + 5% milk for 16 hours at 4°C.
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All lanes : Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324) at 1 µg/ml
Lane 1 : Isolated mitochondria from Human heart at 5 µg
Lane 2 : Isolated mitochondria from Bovine heart at 1 µg
Lane 3 : Isolated mitochondria from Rat heart at 10 µg
Lane 4 : Isolated mitochondria from Mouse heart at 10 µg
Lane 5 : Isolated mitochondria from HepG2 cells at 20 µg
Predicted band size: 22 kDa -
Flow cytometric analysis using ab110324 at 1µg/ml staining Cyclophilin F in HL60 cells (blue). Isotype control antibody (red).
Datasheets and documents
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SDS download
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Datasheet download
References (85)
ab110324 has been referenced in 85 publications.
- Amanakis G et al. Cysteine 202 of cyclophilin D is a site of multiple post-translational modifications and plays a role in cardioprotection. Cardiovasc Res 117:212-223 (2021). PubMed: 32129829
- Portz P & Lee MK Changes in Drp1 Function and Mitochondrial Morphology Are Associated with the a-Synuclein Pathology in a Transgenic Mouse Model of Parkinson's Disease. Cells 10:N/A (2021). PubMed: 33924585
- Li A et al. Butyrate Feeding Reverses CypD-Related Mitoflash Phenotypes in Mouse Myofibers. Int J Mol Sci 22:N/A (2021). PubMed: 34299032
- Dhingra R et al. Impaired NF-?B signalling underlies cyclophilin D-mediated mitochondrial permeability transition pore opening in doxorubicin cardiomyopathy. Cardiovasc Res 116:1161-1174 (2020). PubMed: 31566215
- Zhang B et al. Mitochondrial dysfunction and endoplasmic reticulum stress in calf hepatocytes are associated with fatty acid-induced ORAI calcium release-activated calcium modulator 1 signaling. J Dairy Sci 103:11945-11956 (2020). PubMed: 32981726