Overview

  • Product name
    Anti-Cenexin1/ODF2 antibody
    See all Cenexin1/ODF2 primary antibodies
  • Description
    Rabbit polyclonal to Cenexin1/ODF2
  • Host species
    Rabbit
  • Specificity
    On the basis of the immunogen sequence, we would expect this antibody to recognise the following isoforms of Cenexin1/ODF2 as listed in Swissprot: Q5BJF6-1 (95 kDa), Q5BJF6-2 (89 kDa), Q5BJF6-3 (93 kDa), and Q5BJF6-4 (95 kDa).
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 750 to the C-terminus of Human Cenexin1/ODF2.

    Read Abcam's proprietary immunogen policy (Peptide available as ab43839.)

  • Positive control
    • This antibody gave a positive signal in Hela whole cell lysate and Human testis tissue lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab43840 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 Use a concentration of 1 µg/ml.
ICC/IF Use at an assay dependent concentration. PubMed: 22096071

Target

  • Function
    Seems to be a major component of sperm tail outer dense fibers (ODF). ODFs are filamentous structures located on the outside of the axoneme in the midpiece and principal piece of the mammalian sperm tail and may help to maintain the passive elastic structures and elastic recoil of the sperm tail. May have a modulating influence on sperm motility. Functions as a general scaffold protein that is specifically localized at the distal/subdistal appendages of mother centrioles. Component of the centrosome matrix required for the localization of PLK1 and NIN to the centrosomes. Required for the formation and/or maintenance of normal CETN1 assembly.
  • Tissue specificity
    Testis-specific (at protein level). Highly expressed in cytoplasm of step 2 round spermatids. Detected in the middle piece and extends to about half the principal piece of the sperm tails.
  • Sequence similarities
    Belongs to the ODF2 family.
  • Post-translational
    modifications
    Tyrosine phosphorylated.
  • Cellular localization
    Cytoplasm > cytoskeleton > centrosome. Cell projection > cilium. Cytoplasm > cytoskeleton > centrosome > centriole. Cytoplasm > cytoskeleton > spindle pole. Localized at the microtubule organizing centers in interphase and spindle poles in mitosis. Localized at the distal/subdistal appendages of mother centrioles.
  • Information by UniProt
  • Database links
  • Alternative names
    • 84 kDa outer dense fiber protein antibody
    • Cancer/testis antigen 134 antibody
    • Cenexin 1 antibody
    • Cenexin antibody
    • Cenexin1 variant 1 antibody
    • CT134 antibody
    • FLJ44866 antibody
    • KKT 4 antibody
    • KKT4 antibody
    • MGC111096 antibody
    • MGC9034 antibody
    • ODF 2 antibody
    • ODF 84 antibody
    • odf2 antibody
    • ODF2/1 antibody
    • ODF2/2 antibody
    • ODF84 antibody
    • ODFP2_HUMAN antibody
    • OTTHUMP00000022274 antibody
    • OTTHUMP00000022275 antibody
    • Outer dense fiber of sperm tail 2 antibody
    • Outer dense fiber of sperm tails 2 antibody
    • Outer dense fiber of sperm tails protein 2 antibody
    • Outer dense fiber of sperm tails, 84 kD antibody
    • Outer dense fiber protein 2 antibody
    • Sperm outer dense fiber major protein 2 antibody
    • Sperm tail structural protein antibody
    • Testis specific autoantigen antibody
    see all

Images

  • ab43840 staining Cenexin1 in the mother centriole of a dividing HeLa cell by ICC/IF. DNA is stained blue, Aurora A is stained red, and Cenexin1 is stained green. Cells were fixed in formaldehyde, permeabilized with 0.1% Triton in PBS, and blocked with 2% BSA for 1 hour at 21°C. The primary antibody was diluted 1/200 and incubated with the sample for 1 hour at 21°C. A FITC-conjugated anti-rabbit IgG H+L antibody was used as the secondary.

    See Abreview

  • All lanes : Anti-Cenexin1/ODF2 antibody (ab43840) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 25 µg
    Lane 2 : Human testis tissue lysate - total protein (ab30257) at 20 µg

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 93 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?


    Exposure time: 20 minutes
  • Immunocytochemistry/ Immunofluorescence analysis of human HCT116 cells labeling Cenexin1/ODF2 with ab43840 at 1/400 dilution. Cells were fixed in formaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Staining with ab43840 at 1/400 was carried out for 1 hour at 22°C in PBS buffer. ab150081, a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed secondary antibody was used at 1/200 dilution. DAPI was used to counterstain.

    See Abreview

  • All lanes : Anti-Cenexin1/ODF2 antibody (ab43840) at 1/500 dilution

    Lane 1 : 25 ug HeLa cell lysate
    Lane 2 : 12.5 ug HeLa cell lysate
    Lane 3 : 25 ug HeLa cell lysate with Human Cenexin1/ODF2 peptide (ab43839) at 4 µg/ml
    Lane 4 : 12.5 ug HeLa cell lysate with Human Cenexin1/ODF2 peptide (ab43839) at 4 µg/ml

    Secondary
    All lanes : Anti-Rabbit IgG HRP-conjugated at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 93 kDa
    Observed band size: ~75 kDa why is the actual band size different from the predicted?


    Exposure time: 4 minutes


    Detection of Cenexin 1 by ab43840 in Western blot of HeLa cell lysates. Please see anonymous abreview for further details.

    See Abreview

References

This product has been referenced in:
  • Watanuki S  et al. Characterization of centriole duplication in human epidermis, Bowen's disease, and squamous cell carcinoma. J Dermatol Sci 91:9-18 (2018). ICC/IF . Read more (PubMed: 29615326) »
  • Aziz K  et al. Mosaic-variegated aneuploidy syndrome mutation or haploinsufficiency in Cep57 impairs tumor suppression. J Clin Invest 128:3517-3534 (2018). Read more (PubMed: 30035751) »
See all 17 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HCT116)
Permeabilization
Yes - 0.5% Triton-X100 in PBS
Specification
HCT116
Fixative
Formaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Jan 16 2017

Question

Hello,
I have attached the images mentioned in my previous letter. They illustrate my concern with this antibody. I will try to answer your specific questions:
1. Are you staining fixed tissues (i.e. paraffin-embedded or frozen) or cells? What is the fixative used and fixation protocol?
I it was tissue fixated in 3.7% paraformaldehyde for 30'.
2. Is there a reason why you used this antibody instead of one of our centrosome markers like Ninein or Gamma tubulin?
This antibody reportedly recognized the protein associated with basal foot of the basal body
Ref1), I needed to resolve this particular structure within the basal body.
3. According to SwissProt, the tissue specificity for this protein is testis-specific (at protein level). We also list HeLa cells as a positive control. Have you tested any of these controls?
I am currently awaiting the results of this experiment and may contact you again once we get them.
4.What had you tried for blocking?
5. What was the dilution of the primary antibody used?
The antibody was used in 1:200 dilution. At this dilution the antibody gives an excellent bright signal with almost no background, but the protein, which it recognizes, is localized most likely within the cilia axoneme, (above the apical cell surface) , and totally absent from the centrosomes, which are localized in the apical surface . It is unlikely that blocking may change this fact.
Reference
1. Cell. 2012 Jan 20;148(1-2):189-200.
Coordinated ciliary beating requires Odf2-mediated polarization of basal bodies via basal feet.
Kunimoto K, Yamazaki Y, Nishida T, Shinohara K, Ishikawa H, Hasegawa T, Okanoue T, Hamada H, Noda T, Tamura A, Tsukita S, Tsukita S.

Read More
Answer

Thank you for your reply and for providing this additional information.
Unfortunately the attachment didn't go through. Could you send it again compiled as 1 PDF, PPT, or DOC file?
Please let me know what the results are for your control experiments. Right now it is difficult for us to determine why the antibody isn't working since the application IHC-P hasn't been tested and the tissue type is not validated. If you could run a positive control WB with HeLa or testis lysate, that would provide us with important information about whether or not the antibody is working.
We look forward to hearing more information about the positive control.

Read More

Answer

Thank you for contacting us. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.
You said that you're staining mouse trachea ciliated cells. Are you staining fixed tissues (i.e. paraffin-embedded or frozen) or cells? What is the fixative used and fixation protocol?
According to SwissProt, the tissue specificity for this protein is testis-specific (at protein level). We also list HeLa cells as a positive control. Have you tested any of these controls?
Was the antibody that worked from the other vendor also to this target protein, or was it to a different protein? Is there a reason why you used this antibody instead of one of our centrosome markers like Ninein or Gamma tubulin?
What had you tried for blocking?
What was the dilution of the primary antibody used?
Should the suggestions not improve the results, please do let me know.
In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.
I hope this information is helpful, and I thank you for your cooperation.

Read More

Answer

Thank you very much for providing that information for me. It will be very helpful to us.

I have just had a look at the status of your order of ab43840 and it has been shipped today so will therefore be with you tomorrow as I said. Please do let me know how you get on with this antibody.

Until then, I wish you all the best with your research.

Read More

Answer

Thank you for confirming those details.

I have now arranged for the new vial ofab43840 to be sent out to you. This is on the order number 1097435(Purchase order number FOCR 303778). This should get to you tomorrow. If you have any problems receiving it please dolet me know.

As mentioned in my previous email, it would be very useful for us to look into the problems you have had with ab72505. To this end, if you could share with me the protocol details and some images of the results I would be very grateful.

If you encounterany further problemswith the new antibody ab43840 please dolet me know. I look forward to receiving your reply.

Read More

Answer

Thank you for contacting us yesterday and sorry for the delay in getting back to you.

I was sorry to hear that youhavebeenhavingdifficulties with the anti-CEP170 antibody(ab43840). We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

As discussed over the phone, I would be happy to arrange forthereplacement antibody you requested ()to be sent to you. Could you please just confirm for me that the delivery address that I have for you is correct:

>Once I have this confirmation I will arrange for the antibody ab43840 to be sent out to you.

I understand the problems you have encountered with this antibody and the optimisation you have had to perform in trying to make it work as expected. If possible, so that I can investigate this case further with thelab, would you be able to give me a little more detail about the experiment carried out? If you wouldn't mind could you please fill out the questionnaire I have attached to this email, including any images you have if possible. This information will also allowme to investigate this case internally and initiate additional testing where necessary.

I look forward to receiving your reply.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jun 22 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Formaldehyde
Permeabilization
Yes - PBS, 0.1% Triton
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Apr 08 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Loading amount
25 µg
Specification
HeLa
Treatment
4 ug/ml immunogen peptide
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Apr 08 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up