Anti-XBP1 antibody (ab37152)

Hello,
I am looking for the antibody recognizing both full and spliced forms of XBP1 in mouse cell line.
I bought antibody from (other antibody company) but it did not detect any signal.
I found that AB37152 and AB37151 products react with mice but product sheets say those antibodies only detect full XBP-1.
However, in published paper using your antibody, your antibodies are able to detect both full and spliced forms for XBP-1.
Could you tell me whether those antibodies can detect full and spliced XBP1 signaling in mice cells?

ANSWER:

Thank you for contacting us.

The correct antibody for detection of both splice forms (as described at the UniProt page for the mouse protein, http://www.uniprot.org/uniprot/O35426 ) is ab37152:

Click here (or use the following: http://www.abcam.com/index.html?datasheet=37152).

This is raised against an N-terminal sequence that is shared by both isoforms.

Antibody ab37151 is raised against a sequence that is found only on the shorter isoform. I do not have an explanation for the result seen in the lower of the two western blots on the datasheet, the blot of HepG2 and placenta. A band at approximately 42 kDa appears but this is unlikely to be the isoform 2, long splice form of XBP1. I have asked the lab to re-test ab37151.

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

I have to contact you again, because of the antibody for XBP1 (ab37152). The problems with this antibody I explained you before. Consequently you sent me a new Aliquot of this antibody (Lot.No.: GR45274-2 ). I repeated my experiment, which I send you before, where I used the old aliquot of ab37152 I have (Lot.No.: 955532). Therefore see also the file attached. To summarize my results: The new antibody (Lot.No.: GR45274-2), which should be specific for both forms the unspliced and spliced XBP1, and the old antibody (Lot.No.: 955532 ) reveal the same reult by only detecting the spliced form of XBP1. Therefore I ask you to send me an aliquot of your u form specific antibody (ab37151) to exclude resp. verify the functionality of ab37152. Thanks a lot in advance

ANSWER:

Thank you for confirming these details and for sending images of your results. I will forward these data to the lab, this will enable us to closely monitor the quality of our products. I am sorry the new lot ab37152 did not perform as stated on the datasheet. As requested, I have issued a free of charge replacement with one unit of ab37151; the order number is *******. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

Would like to test ab37152 with rat samples.

ANSWER:

Thank you for your call today and for your questions. As we discussed, I would be happy to help resolve the issues with the antibodies that you have purchased. Could you please send the following information about the protocol and results using ab79483, ab11419, and ab50354: 1) How was the lysate prepared, and how much of the rat brain lysate was loaded per lane of the gel? 2) What kind of blocking solution was used? 3) What dilution of antibody was used? 4) Have the secondary antibodies and detection systems been working with other primary antibodies? 5) Do you have any images that you could send for our reference? If non-specific bands were seen on theblots, were any of them more predominant than the others? I look forward to hearing from you and resolving these issues. Please see below for information about your testing discount code for ab37152. DISCOUNT CODE: ***Expiration date: May 13, 2012 This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview forab37152 in ratand include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. 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.

I think that's a good idea to receive a free unit of the new lot ab37152 to test it against both unspliced and spliced XBP1. T

ANSWER:

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. 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. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

To whom it may concern, I'm currently working on the protein XBP1, therefore I ordered your antibody ab37152, which should detect both forms of XBP1 (unspliced and spliced form). In different experimental approaches I tried to detect the unspliced form of XBP1 via wetsern blot. Therefore I overexpressed both the unspliced and the spliced form of XBP1 with respective expression plasmids in human cells. Additionally the degradation via proteasome and the splicing of the u-form by IRE1 were inhibited (Please see attachment). To summarize: I overexpressed the u-form of XBP1 and inhibited the conversion to the s-form and the degradation of the protein and could not detect the u-form with the XBP1 antibody ab37152. Therefore I would like to purchase the antibody only specific for the u-form of XBP ab37151. In the case that the antibody which should detect both forms, even detects the spliced form of XBP1, I would be very pleased, if you could give me an appropriate offer for this antibody. It would be also possible to send me a small aliquot of the antibody (u-specific, ab37151) to test it for my application. About a further troobleshooting concerning the detection of the unspliced form with antibody ab37152 I would be very pleased as well. If there are any questions left, do not hesitate to contact me. --

ANSWER:

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results. The quality of our products is very important for us, and I would therefore like to investigate this issue further. Please see below. Indeed, if we do conclude that the antibody is not working as stated on the datasheet, we will replace or refund it to you (if purchased in the last 6 months). In regards ab37151: we do not have small sized vials for testing purposes. I am sorry. We do however have again the guarantee on our products. Furthermore, should you need to use this antibody on an untested species/application, I might can provide a testing discount. If you use the antibody on a tested application, we invite you also to submit product reviews (Abreviews) directly on the respective datasheet. This will provide you with generous points which can be used for discounts or amazon vouchers. As said above, I would like to review the problem with the ab37152. Can you therefore please provide we with more information, such as what sequences, of what species, have been overexpressed - what controls have been used? I have also attached a questionnaire to this email, which will help to put the necessary information together. It should not take you more than 10 minutes to fill it all out. I am looking forward to hear back from you, and I thank you for your cooperation.