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  1. Link

    cyclophilin-f-antibody-epr11311-121-ab231155.pdf

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Cell Biology Apoptosis Mitochondrial
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
  • Flow Cytometry (Intracellular) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
  • Western blot - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
  • Western blot - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
  • 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)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
  • Flow Cytometry (Intracellular) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
  • Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)

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

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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: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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 Cyt (intra): 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR11311-121
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Mitochondrial
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Mitochondrial
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism

Associated products

  • Alternative Versions

    • Anti-Cyclophilin F antibody [EPR11311-121] - BSA and Azide free (ab231287)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Immunohistochemistry reagents

    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human PPIF (Cyclophilin F) knockout HEK-293T cell line (ab266077)
  • KO cell lysates

    • Human PPIF (Cyclophilin F) knockout HEK-293T cell lysate (ab257039)
  • KO cell pellets

    • Human PPIF (Cyclophilin F) knockout HEK-293T cell pellet (ab279017)
  • Recombinant Protein

    • Recombinant human Cyclophilin F protein (ab79186)
  • Related Products

    • Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)
    • Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/600.
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.
Notes
Flow Cyt (Intra)
1/600.
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.

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.
  • Target information above from: UniProt accession P30405 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10105 Human
    • Entrez Gene: 105675 Mouse
    • Entrez Gene: 282819 Rat
    • Omim: 604486 Human
    • SwissProt: P30405 Human
    • SwissProt: Q99KR7 Mouse
    • SwissProt: P29117 Rat
    • Unigene: 381072 Human
    • Unigene: 41656 Mouse
    • Unigene: 2923 Rat
    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 (Intracellular) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)
    Flow Cytometry (Intracellular) - Anti-Cyclophilin F antibody [EPR11311-121] (ab231155)

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

    Intracellular 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

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (1)

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

ab231155 has been referenced in 1 publication.

  • Yu CH  et al. TDP-43 Triggers Mitochondrial DNA Release via mPTP to Activate cGAS/STING in ALS. Cell 183:636-649.e18 (2020). PubMed: 33031745

Customer reviews and Q&As

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