Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)

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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: WB, IHC-P, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Cyclophilin F antibody [EPR11311-121]
    See all Cyclophilin F primary antibodies

  • Description

    Rabbit monoclonal [EPR11311-121] to Cyclophilin F

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Cyclophilin F aa 1-100. The exact sequence is proprietary.
    Database link: P30405

  • Positive control

    • 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 Cytometry: HeLa and Jurkat cells.

  • General notes

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab231155 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
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.
Flow Cyt 1/600.

Target

  • 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

  • Database links

    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
    • Cyp D antibody
    • CyP M antibody
    • CyP-D antibody
    • CyP-M antibody
    • CYP3 antibody
    • CypD antibody
    • hCyP3 antibody
    • mitochondrial antibody
    • Mitochondrial cyclophilin antibody
    • Peptidyl prolyl cis trans isomerase, mitochondral antibody
    • Peptidyl-prolyl cis-trans isomerase F antibody
    • Peptidyl-prolyl cis-trans isomerase F, mitochondrial antibody
    • Peptidylprolyl isomerase F (cyclophilin F) antibody
    • Peptidylprolyl isomerase F antibody
    • PPIase antibody
    • PPIase F antibody
    • Ppif antibody
    • PPIF_HUMAN antibody
    • Rotamase antibody
    • Rotamase F antibody
    see all

Images

  • Western blot - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    Western blot - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)

    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).

  • Flow Cytometry - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    Flow Cytometry - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)

    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 (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.

  • Western blot - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    Western blot - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    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 seconds


    ab231155 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.

  • Western blot - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    Western blot - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    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 

  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)

    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).

  • Flow Cytometry - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    Flow Cytometry - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)

    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.

  • Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)

Protocols

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.

Click here to view the general protocols

References (0)

Publishing research using ab231155? Please let us know so that we can cite the reference in this datasheet.

ab231155 has not yet been referenced specifically in any publications.

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