Anti-Citrulline antibody (ab100932)

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.

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.

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.

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.

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.

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.

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.

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.

Thank you for your reply.

The Abpromise guarantee is only valid for products bought direct from Abcam or one of our authorized distributors. The terms and conditions of the guarantee can be found on https://www.abcam.com/abpromise.

I look forward to your reply with the order details.

Thank you for your reply.

As a compensation I can send to you one unit of ab6464, unfortunately I cannot send both ab6464 and ab100932.

There is no order in our system corresponding to the address you provided in your last email. Could you please send me an order number as well as the date the order was placed? Have you ordered the product directly at Abcam orvia a subdealer?

I look forward to your reply.