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Anti-INCENP antibody (ab12183)

Reviews (6)Q&A (6)References (20)

Key features and details

  • Rabbit polyclonal to INCENP
  • Suitable for: IP, ICC, WB, ICC/IF, IHC-Fr
  • Reacts with: Mouse, Rat, Human, Xenopus laevis
  • Isotype: IgG

Overview

  • Product name

    Anti-INCENP antibody
    See all INCENP primary antibodies

  • Description

    Rabbit polyclonal to INCENP

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, ICC, WB, ICC/IF, IHC-Frmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human, Xenopus laevis
    Predicted to work with: Chicken

  • Immunogen

    Synthetic peptide:

    SKPRYHKRTSSAVWNSP

    (with N-terminal added cysteine) conjugated to KLH, corresponding to amino acids 884-901 of Human INCENP.
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • General notes

    If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilutions should be discarded if not used within 12 hours. There is some debate about whether this antibody is suitable for use in IF and we would be most interested to hear from researchers who test this antibody in this application. The antibody has been reported to be suitable for use in IF at a dilution of 1/100 using HeLa cells (see image below). However, one of our customers used this product in HeLa cells and reported an unusual staining pattern - please see the enquiries section of the online datasheet for further details.

    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 ab12183 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
IP
Use at an assay dependent concentration.
ICC (2)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.
ICC/IF (3)
Use at an assay dependent concentration.
IHC-Fr (1)
Use at an assay dependent concentration.
Notes
IP
Use at an assay dependent concentration.
ICC
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.

Target

  • Function

    Component of the chromosomal passenger complex (CPC), a complex that acts as a key regulator of mitosis. The CPC complex has essential functions at the centromere in ensuring correct chromosome alignment and segregation and is required for chromatin-induced microtubule stabilization and spindle assembly. Probably acts through association with AURKB or AURKC. Seems to bind directly to microtubules. Controls the kinetochore localization of BUB1.

  • Sequence similarities

    Belongs to the INCENP family.

  • Cellular localization

    Chromosome > centromere. Cytoplasm > cytoskeleton > spindle. Nucleus. Chromosome > centromere > kinetochore. Localizes to inner kinetochore. Localizes on chromosome arms and inner centromeres from prophase through metaphase and then transferring to the spindle midzone and midbody from anaphase through cytokinesis. Colocalizes with AURKB at mitotic chromosomes.
  • Information by UniProt

  • Database links

    • Alternative names

      • binds and activates aurora B and C in vivo and in vitro antibody
      • Chromosomal passenger protein antibody
      • INCE_HUMAN antibody
      • INCENP antibody
      • Inner centromere protein antibody
      • Inner centromere protein antigens 135/155kDa antibody
      • Inner centromere protein antigens 135kD 155kD antibody
      • Inner centromere protein INCENP antibody
      see all

    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

    References (20)

    Publishing research using ab12183? Please let us know so that we can cite the reference in this datasheet.

    ab12183 has been referenced in 20 publications.

    • Park M  et al. A novel synthetic microtubule inhibitor exerts antiproliferative effects in multidrug resistant cancer cells and cancer stem cells. Sci Rep 11:10822 (2021). PubMed: 34031528
    • Tsunematsu T  et al. APC/CCdh1 is required for the termination of chromosomal passenger complex activity upon mitotic exit. J Cell Sci 133:N/A (2020). PubMed: 32934012
    • Kim S  et al. PRMT6-mediated H3R2me2a guides Aurora B to chromosome arms for proper chromosome segregation. Nat Commun 11:612 (2020). PubMed: 32001712
    • Abad MA  et al. Borealin-nucleosome interaction secures chromosome association of the chromosomal passenger complex. J Cell Biol 218:3912-3925 (2019). PubMed: 31570499
    • Konishi M  et al. Quantitative analyses of the metaphase-to-anaphase transition reveal differential kinetic regulation for securin and cyclin B1. Biomed Res 39:75-85 (2018). PubMed: 29669986
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a question

    1-6 of 6 Q&A

    Question

    I would like to see the western blot used to confirm that this antibody recognises INCENP. The data sheet states that the predicted size of INCENP is 106 Kda, but does not say whether the antibody recognises a band of this size on a western blot?? I find the antibody recognises a single band of 80kDa on whole cell HeLa lysate. I see you have no independent reviews of this antibody and the technical inquiries increase my doubt that this antibody recognises INCENP. It seems unwise to continue to market this antibody as "Anti-INCENP" without better (any?) evidence that it recognises INCENP. I would appreciate any data you have on the performance of this antibody in order that I can gain something from the experiments where I have used it. It may be, for some reason, that the 80kDa band I see is INCENP, running at an unusual size. Currently I am unsure what it is! I realise that I am too late for a refund, but that is of little concern compared to the time that I and others may waste from an antibody that recognises the wrong protein! Thanks for your help,

    Read More
    Answer

    I have received confirmation from the source of ab12183 that the antibody will detect a 106 kDa band using whole HeLa extract. They cannot explain why you are detecting a 80kDa band and hypothesize that this could be a splice variant or a lysed form of INCENP. Unfortunately they were unable to provide us with an image of the blot, I apologise for the inconvenience,

    Read More

    Question

    BATCH NUMBER 63014 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM I have some doubt about whether this antibody recognises INCENP in a western blot. When used against total HeLa lysate it gives a single band of 80kDa. This band fails to disappear in lysates of siRNAi-treated HeLa, in which INCENP appears to be knocked down by western and IF with another INCENP antibody. I would be interested to hear how it has been confirmed that this antibody recognises INCENP. HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? >10 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

    Read More
    Answer

    We have just received the following information from the source of ab12183 who tested the antibody in HeLa nuclear extracts: 1. Extract: 10^7 cells/slab of HeLa nuclear. 2.Wet transfer for 90 minutes at 4'C. 3. Blocking solution: 5%-10% non fat dry milk for 2-24 hours at room temperature. 4. Wash and Dilution buffer: 1% BSA in PBS-T. 5. Primary antibody: this product at 1:5,000-1:10,000 dilutions. Incubation for 120 minutes at room temperature. 6. Secondary antibody: HRP anti rabbit IgG at 1:20,000 dilution. Incubation for 60 minutes at room temperature. 7. Substrate: ECL. I hope this information helps, please do not hesitate to contact us if you need further assistance,

    Read More

    Question

    We have tried all of your suggestions and nothing is working. You suggested: - "First, I suggest running a positive control." our response - IVP is a positive control - "you did not mention what dilutions you have tried, but you may need to increase the concentration of the primary. Use a minimum dilution of 1/2000 (recommended when using a whole extract of mouse NIH-3T3 and rat PC12 cells and a chemiluminescent detection reagent)- 1/5000 (recommended when using a nuclear extract of human HeLa cells and a chemiluminescent detection reagent)." our response - we used a dilution of 1/1000 - You may also need to increase the concentration of your secondary antibody, and extend the incubation periods for both the primary and secondary. For example, incubate overnight at 4C with ab12183. our response - we icubated at 4C overnight. We would like to return the antibody and have it replaced with AB2270.

    Read More
    Answer

    Thanks for your email. I can send you ab2270 as a replacement. There's no need to return ab12183. Can you please confirm your shipping address, including phone number?

    Read More

    Question

    BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER Credit C DESCRIPTION OF THE PROBLEM No signal or weak signal, no band (not recognizing protein) SAMPLE IVT and IP PRIMARY ANTIBODY ab12183 (INCENP antibody) SECONDARY ANTIBODY Donkey anti-rabbit DETECTION METHOD ECL+ POSITIVE AND NEGATIVE CONTROLS USED IVT=positive control ANTIBODY STORAGE CONDITIONS 4? SAMPLE PREPARATION (MP40) / cocktail / 95? C for 10 minutes AMOUNT OF PROTEIN LOADED 1microlitre of IVT product ELECTROPHORESIS/GEL CONDITIONS SDS-PAGE, 15% TRANSFER AND BLOCKING CONDITIONS Tris + SDS + METH / 200 MA for 1 hour and 15 minutes, 4% milk in TBST HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? No ADDITIONAL NOTES verified the presence of IVT product by phosphoImager using S35

    Read More
    Answer

    Thank you for your enquiry, and I'm sorry to hear that you are experiencing difficulty with ab12183. I would like to make some suggestions in order to assist you. First, I suggest running a positive control. For this antibody nuclear/whole extract of human HeLa cells is recommended. Also, you did not mention what dilutions you have tried, but you may need to increase the concentration of the primary. Use a minimum dilution of 1/2000 (recommended when using a whole extract of mouse NIH-3T3 and rat PC12 cells and a chemiluminescent detection reagent)- 1/5000 (recommended when using a nuclear extract of human HeLa cells and a chemiluminescent detection reagent). You may also need to increase the concentration of your secondary antibody, and extend the incubation periods for both the primary and secondary. For example, incubate overnight at 4C with ab12183. If you have any additional questions, please contact us again.

    Read More

    Question

    I have tried to use this antibody to stain Hela cells(methanol fixation). It stained the centrosomes clearly at all stages of cell cycle. So far, INCENP is recognized as chromosomal passenger protein. It should localize to the centromere around metaphse, spindle midzone during anaphase. So I don't think this antibody could be used for immunostaining. If you company cann't repeat the supplier's claim, we will have to return the reagent to your company.

    Read More
    Answer

    Thank you for getting back to me with an explanation of the staining you would expect with this antibody. I will remove the recommendation that it is suitable for use in IF and refund you for the cost of the vial you purchased from us. (Order number 49851 shipped on 16 August 2004). Thank you again for your feedback.

    Read More

    Question

    I bought the antibody ab12183 and did immunostaining using Hela cells. Unexpectedly, this antibody only stained the centrosomes, not the typical chromosomal passenger proteins staining. How could this happen?

    Read More
    Answer

    Chromosomal passenger proteins are found in different places in the cell depending on the stage of the cell cycle - please see the relevance section of the datasheet for more details. Your message suggests that the antibody should not stain the centrosomes, and I would be interested to know why you think that centrosomes should not be stained. We are trying to reproduce the results of our supplier who claims it can be used in IF on HeLa cells at a 1:100 dilution, but we have not been able to obtain a positive result (U2OS cells were used in our experiments). I'd be very interested to know more about your experience with INCENP and to work together to confirm whether this antibody is suitable for use in IF.

    Read More

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