Overview

  • Product name

  • Description

    Rabbit polyclonal to Citrulline
  • Host species

    Rabbit
  • Specificity

    ab100932 reacts specifically with intrapeptidic citrulline independently of the amino acid sequence. It does not react with free citrulline, ornithine or arginine. Citrulline is an un-natural amino-acid and its expression is very low. Factors limiting the detection are the amount of endogenous citrulline within a protein and the accessibility of the antibody to these sites.
  • Tested applications

    Suitable for: ELISA, ICCmore details
    Unsuitable for: WB
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Citrulline coupled to KLH via Glutaraldehyde

  • Positive control

    • Hypercitrullinated Histones induced in HL60 cells by DMSO +calcium +calcium ionophore A23187

Properties

Applications

Our Abpromise guarantee covers the use of ab100932 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
ELISA Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for WB.
  • Target

    • Relevance

      The amino acid Citrulline is required to detoxify the liver from ammonia, which is a waste product of the body from oxidation. Citrulline promotes energy and assists with the immune system. This unusual amino acid is formed in the urea cycle by the addition of carbon dioxide and ammonia to ornithine. It is then combined with aspartic acid to form arginosuccinic acid, which later is metabolized into the amino acid arginine.

    Images

    • ELISA data showing the high immuno-reactivity of ab100932 against citrulline due to the high amounts of citrulline conjugated to BSA.

    • ELISA data demonstrating the specificity of ab100932 to citrulline naturally present in histones (endogenous citrulline).

    • Immunocytochemical analysis of Human buccal mucosal cells, staining Citrulline with ab100932.

      Cells were fixed with acetone followed by 2% glutaraldehyde, permeabilized with 0.1% Triton X-100 and blocked with 3% BSA for 20 minutes at 22°C. Samples were incubated with primary antibody (1/200 in PBS) for 15 hours at 4°C. An undiluted biotinylated anti-rabbit polyclonal IgG was used as the secondary antibody.

      See Abreview

    References

    This product has been referenced in:

    • Verheul MK  et al. Pitfalls in the detection of citrullination and carbamylation. Autoimmun Rev 17:136-141 (2018). Read more (PubMed: 29203292) »
    • Hutchinson D  et al. Cadmium nanoparticles citrullinate cytokeratins within lung epithelial cells: cadmium as a potential cause of citrullination in chronic obstructive pulmonary disease. Int J Chron Obstruct Pulmon Dis 13:441-449 (2018). Read more (PubMed: 29430177) »
    See all 9 Publications for this product

    Customer reviews and Q&As

    1-10 of 18 Abreviews or Q&A

    This product is known to not work in this application or species.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa P4, HEK293FRT)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    10 µg
    Specification
    HeLa P4, HEK293FRT
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Mrs. Saskia Hutten

    Verified customer

    Submitted Oct 05 2018

    Question
    Answer

    Ab100932 was tested on ELISA on various proteins and peptides containing citrulline and gave positive results. An inhibition with free citrulline, ornithine or arginine was also performed and show no decrease of the signal on citrullinated protein or peptide.

    Read More

    Answer

    WB can be tried using dilution between 1/500- 1/1000.
    ELISA can tried using 1/500 dilution.
    The information about positive control is given in Hagiwara et. al. 2001.

    Read More
    Application
    Immunocytochemistry
    Sample
    Human Cell (buccal mucosa)
    Specification
    buccal mucosa
    Fixative
    acetone followed by 2% gluteraldehyde
    Permeabilization
    Yes - 0.1 % triton in blocking buffer
    Blocking step
    BSA as blocking agent for 20 minute(s) · Concentration: 3% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted Jan 29 2013

    Answer

    I am sorry to hear that this antibody is not providing satisfactory results.

    Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab100932. I would also appreciate if you can confirm some further details:

    Typically a high background with white bands or "ghost bands" indicates a signal that is too strong. You can reduce the primary antibody even further and/or add additional washes after primary incubation. Another option is to incubate only 1 hr at room temperature with the primary. You may also want to switch to 5% milk for blocking, at times when the BSA is giving high background sometimes switching the blocking agent solves the problem.

    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

    Question
    Answer

    DISCOUNT CODE: ***

    Expiration date: January 28th, 2013

    I am very pleased to hear you would like to accept our offer and test ab100932 inIHC. This code will give you: 1 freeprimary antibodybefore the expiration date. To redeem this offer, please submit an Abreview forIHC and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
    For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.

    Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

    Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

    The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

    Read More

    Question
    Answer

    Thank you for your inquiry.

    The lab did not have a positive control testing WB image, however, they were able to pass along the protocol tested:

    The positive control was HL60 cells treated with DMSO or with DMSO and A23187. The histones of these cells were then extracted.
    The sample were loaded on a 14% acrylamide SDS PAGE gel.
    The transfer was made on a PVDF membrane
    The blocking buffer was PBS 1X with 5% BSA
    The antibody was used at 1/1000 in a TBST Tween 20 buffer with 1% BSA.
    The antibody was incubated overnight at +4°C.

    As I mentioned on the phone, the 4 for 3 promotion runs through the end of this month. If you buy 4 products, the least expensive one is free! Just mention 4FOR3-TB3ME on your online or phone order. I hope this information helps. Please contact us with any other questions.

    Read More

    Answer

    Thank you for contacting us. We currently have two antibodies that will detect the citrulline modification on any proteins: ab6464 and ab100932. While ab6464 has been validated in IHC on frozen tissue sections, we have not tested this antibody for use in WB. Similarly, ab100932 has been validated in WB but not in IHC. If you would like to test either of these antibodies, I am happy to offer you our 100% testing discount program. If you purchase the antibody for full price and submit an Abreview with your protocol and results using the antibody in an untested application, you can get a discount code for a free primary antibody on a future purchase. Whether or not the antibody works, the next antibody you buy from us would be free. If you are interested in this program, please reply and let me know which antibody you would like to test, and I will be happy to set up the code for you.

    Read More

    Answer

    Thank you for confirming these details and for your cooperation.

    As requested, I have issued a free of charge replacement with the order number *******.

    To check the status of the order please contact our Customer Service team and reference this number.

    Read More

    Answer

    Thank you for confirming these details and for your cooperation.

    As agreed, I have issued a free of charge replacement with one unit of ab6464,the order number ******.

    To check the status of the order please contact our Customer Service team and reference this number.

    I wish you the best of luck with your research.

    Read More

    1-10 of 18 Abreviews or Q&A

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