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Validated using a knockout cell line

Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)

Reviews (4)Q&A (8)References (85)
Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
  • Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
  • Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
  • Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
  • Flow Cytometry - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)

Key features and details

  • Mouse monoclonal [E11AE12BD4] to Cyclophilin F
  • Suitable for: WB, Flow Cyt, ICC/IF, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Cow, Human
  • Isotype: IgG1

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488

Overview

  • Product name

    Anti-Cyclophilin F antibody [E11AE12BD4]
    See all Cyclophilin F primary antibodies

  • Description

    Mouse monoclonal [E11AE12BD4] to Cyclophilin F

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IF, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Human

  • Immunogen

    Recombinant full length protein corresponding to Rat Cyclophilin F aa 1-206. (also known as CypD)

  • Positive control

    • Isolated mitochondria from Human heart, Bovine heart, Rat heart, Mouse heart, HEK293T, HepG2 cells; Cultured Human embryonic lung-derived fibroblasts (strain MRC5); Human cerebellum tissue; HL60 cells.

  • General notes

    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.

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
WB (2)
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.
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.
Notes
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.
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.

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 [E11AE12BD4] (ab110324)
    Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)

    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/ Immunofluorescence - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
    Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
    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).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
    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.
  • Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
    Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
    All lanes : Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324) at 1/10000 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 - ab110324 observed at 23 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

     ab110324 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. ab110324 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
    Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)This image is courtesy of an anonymous Abreview
    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

    Secondary
    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.

    See Abreview

  • Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
    Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
    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 Cytometry - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
    Flow Cytometry - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)
    Flow cytometric analysis using ab110324 at 1µg/ml staining Cyclophilin F in HL60 cells (blue). Isotype control antibody (red).

References (85)

Publishing research using ab110324? Please let us know so that we can cite the reference in this datasheet.

ab110324 has been referenced in 85 publications.

  • Amanakis G  et al. Cysteine 202 of cyclophilin D is a site of multiple post-translational modifications and plays a role in cardioprotection. Cardiovasc Res 117:212-223 (2021). PubMed: 32129829
  • Portz P & Lee MK Changes in Drp1 Function and Mitochondrial Morphology Are Associated with the a-Synuclein Pathology in a Transgenic Mouse Model of Parkinson's Disease. Cells 10:N/A (2021). PubMed: 33924585
  • Li A  et al. Butyrate Feeding Reverses CypD-Related Mitoflash Phenotypes in Mouse Myofibers. Int J Mol Sci 22:N/A (2021). PubMed: 34299032
  • Dhingra R  et al. Impaired NF-?B signalling underlies cyclophilin D-mediated mitochondrial permeability transition pore opening in doxorubicin cardiomyopathy. Cardiovasc Res 116:1161-1174 (2020). PubMed: 31566215
  • Zhang B  et al. Mitochondrial dysfunction and endoplasmic reticulum stress in calf hepatocytes are associated with fatty acid-induced ORAI calcium release-activated calcium modulator 1 signaling. J Dairy Sci 103:11945-11956 (2020). PubMed: 32981726
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-10 of 12 Abreviews or Q&A

Western blot abreview for Anti-Cyclophilin F antibody [E11AE12BD4]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (kidney)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 23 2021

Immunohistochemistry (Frozen sections) abreview for Anti-Cyclophilin F antibody [E11AE12BD4]

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Heart)
Permeabilization
Yes - Triton x-100, 0.03%
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 30 2017

Western blot abreview for Anti-Cyclophilin F antibody [E11AE12BD4]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - other (Mouse liver mitochondria)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
25 µg
Specification
Mouse liver mitochondria
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 01 2015

Immunoprecipitation abreview for Anti-Cyclophilin F antibody [E11AE12BD4]

 Good
Abreviews
abreview image
Application
Immunoprecipitation
Immuno-precipitation step
Protein A/G
Sample
Mouse Tissue lysate - whole (brain)
Specification
brain
Total protein in input
10 µg
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 05 2014

Question

Je souhaiterais savoir si vous aviez des informations de cross-réactivité avec les Cyclophilines A, B et C pour les anticorps anti-Cyclophiline F suivants :

- ab86425

- ab64935

- ab3567

- ab37664

- ab110324

Read More
Answer


Sachez que ces anticorps sont contre la cyclophiline F ', PPIF, CYP3, SwissProt: P30405. Toutefois, dans le domaine mitochondrial, cette protéine est généralement appelé CYPD ou cyclophiline D - une protéine de 25 kDa dans les mitochondries.
Je peux vous confirmer qu’ab110324, ab64935 et ab3567 ne croisent pas avec les Cyclophilines A, B et C. Cependant, pour ab86425 et ab37664 nous n’avons pas déterminé si ces anticorps croisent avec les Cyclophilines A, B et C.

Read More

Question

Customer odered this product as she thought it is the mitoch. expressed protein. However, there are two proteins labbeled as Cyclophilin D, and this abs detects the cytoplasmatic form. Unfortunately, it's not very clear on the ds. The same is vice versa for the mitochondrial. This info can be found, but WAY down on the ds.
Worse, the ds for the cytoplasmatic abs, ab135717 had been changed in Oct 2012.
Customer is getting a FOCR

Read More
Answer

Vielen Dank für Ihren Anruf.

Es tut mir leid, dass Sie Probleme mit diesem Antikörper hatten, nur weil wir einen Datenblattfehler gemacht haben. Wie am Telefon besprochen habe ich eine kostenlose Ersatzlieferung für Sie in Auftrag gegeben. Sie hat die Referenznummer 123456 und sollte Dienstag bei Ihnen ankommen.

Ich wünsche Ihnen viel Erfolg für Ihr Projekt. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

Read More

Question

Inquiry: Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324) Does this antibody recognize Cyclophilin A? We want to only look at cyclophilin D.

Read More
Answer

Thank you for contacting us.



The antibody was raised against full length recombinant rat CypD protein.

The identity between cyp A and “cypD” (cylophilin F, PPIF) is only 75% which is not high.

The antibody shows no bands in CypD knockouts reported in the literature, also blotting with mitochondrial fractions and immunofluorescent microscopy indicate only a mitochondrial location.

Therefore, we feel vert confident in the specificity of this antibody.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Question

J’utilise votre anticorps anti-cyclophilin F (ab110324) et j’aimerais connaître son épitope ou au moins sa localisation (n ou c-term) svp.

Read More
Answer



L'anti-cyclophilin F ab110324 a été développé en utilisant la protéine entière et malheureusement nous ne disposonsd'aucune information sur l'épitope, ni même la région reconnue.

Read More

Question

Hi Abcam,

A customer would like to know what the predicted band size is for this antibody.

On the image provided on the datasheet the predicted band size is 41kDa, but this looks more like 22 kDa.

Could you please confirm the above?

Kind regards,

Read More
Answer

Thank you for your enquiry.

I have investigated the details for ab110324 Cyclophilin F antibody,I can confirm that the correct predicted molecular weight, according to the WB data on the datasheet and the SwisProt/Uniprot protein database link for Cyclophilin F is 22 kDa.

The datasheet has been updated with the correct information and I apologize for any confusion.

The quality of our antibodies and datasheets is very important to us, as is any feedback and data we receive. It is regrettable that the information we had on the datasheet forthis particular antibody was not of the standard both we and our customers expect.

Should you have any further questions regarding this issue, please do not hesitate to contact me.

Read More

Question

We've retried the antibody and it seems to be working just fine now - not
sure what the problem initially but it seems to of been at our end.
Thanks for your quick responses and follow ups - very much appreciated.

Read More
Answer

Thank you for your reply. I am glad to hear that the antibody is working well for you now. Please let me know if you have any further questions or concerns.

Read More

1-10 of 12 Abreviews or Q&A

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