Validated using a knockout cell line
Recombinant
RabMAb

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

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

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

    This product is a recombinant rabbit monoclonal antibody.

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

  • 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

  • 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.

  • 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.

  • 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 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.

  • 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 

  • 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.

  • 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).

  • 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).

  • 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 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.

References

ab231155 has not yet been referenced specifically in any publications.

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