Product nameAnti-Cyclophilin F antibody [E11AE12BD4]
See all Cyclophilin F primary antibodies
DescriptionMouse monoclonal [E11AE12BD4] to Cyclophilin F
Tested applicationsSuitable for: WB, Flow Cyt, In-Cell ELISA, ICC/IF, IHC-P, IPmore details
Species reactivityReacts with: Mouse, Rat, Cow, Human
Recombinant full length protein corresponding to Rat Cyclophilin F aa 1-206. (also known as CypD)
- Isolated mitochondria from Human heart, Bovine heart, Rat heart, Mouse heart, HepG2 cells; Cultured Human embryonic lung-derived fibroblasts (strain MRC5); Human cerebellum tissue; HL60 cells.
This antibody clone is manufactured by Abcam.
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.
Storage instructionsShipped at 4°C. Store at +4°C. Do Not Freeze.
Storage bufferPreservative: 0.02% Sodium azide
Constituent: HEPES buffered saline
Concentration information loading...
Purification notesab110324 was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation.
Light chain typekappa
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.
|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.
|In-Cell ELISA||Use a concentration of 4 µg/ml.|
|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.|
FunctionPPIases accelerate the folding of proteins. It catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides.
Sequence similaritiesBelongs to the cyclophilin-type PPIase family.
Contains 1 PPIase cyclophilin-type domain.
Cellular localizationMitochondrion matrix.
- Information by UniProt
FormThis 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.
- Cyclophilin 3 antibody
- cyclophilin D antibody
- Cyclophilin F antibody
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.
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).
All 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
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 seconds
Blocked 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.
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.
Immunocytochemistry/ Immunofluorescence analysis of HEK293 cells labeling Cyclophilin F with ab110324 at 1/200 dilution. Cells were fixed with paraformaldehyde and permeabilized with 1% triton x-100. 10% goat serum was used to blocke the cells for 1 hour at room temp followed by incubation with Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324) in 10% goat serum-PBST for 16 hours at 4°C. A goat anti-mouse IgG secondary antibody was used at 1/300 dilution.
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).
This product has been referenced in:
- Xu T et al. ARC regulates programmed necrosis and myocardial ischemia/reperfusion injury through the inhibition of mPTP opening. Redox Biol 20:414-426 (2019). Read more (PubMed: 30415165) »
- Shivram H et al. MicroRNAs reinforce repression of PRC2 transcriptional targets independently and through a feed-forward regulatory network. Genome Res 29:184-192 (2019). Read more (PubMed: 30651280) »