Overview

  • Product name
    Anti-Cyclophilin F antibody
    See all Cyclophilin F primary antibodies
  • Description
    Rabbit polyclonal to Cyclophilin F
  • Tested applications
    Suitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Chimpanzee, Orangutan
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Cyclophilin F.

  • Positive control
    • WB: Human heart and heart mitochondrial tissue lysates and HeLa, HepG2, HEK293 and HL60 whole cell lysates. ICC/IF: A431 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab126573 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).

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
    • 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



  • Predicted band size : 22 kDa

    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 - ab126573 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab126573 was shown to recognize Cyclophilin F when Cyclophilin F knockout samples were used, along with additional cross-reactive bands. Wild-type and Cyclophilin F knockout samples were subjected to SDS-PAGE.  Ab126573 and ab8245 (Mouse 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. 

  • ICC/IF image of ab126573 stained A431 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab126573, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • All lanes : Anti-Cyclophilin F antibody (ab126573) at 1 µg/ml

    Lane 1 : Human Heart Mitochondrial Lysate
    Lane 2 : Human heart tissue lysate - total protein (ab29431)
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate
    Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 6 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 22 kDa
    Observed band size : 22 kDa
    Additional bands at : 48 kDa,61 kDa,96 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 30 secondsThis blot was produced using a 10% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab126573 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

References

ab126573 has not yet been referenced specifically in any publications.

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