Anti-ENPP2/ATX抗体[1F8] (ab77104)

Based on the immunogen for ab77104, the antibody should recognize all isoforms, but we have not specifically tested this. The immunogen is the full length ENPP2 protein.

Thank you for contacting Abcam.

I've compiled a list of primary antibodies that have been tested in mouse tissue via western blot. The only ovary-specific antibodies that I found were anti-human (with only 50% homology with the respective mouse protein), so the ovary antibody listed below is not exclusively-specific to ovary but also gives a positive signal in kidney, placenta, and small intestine:

1. Liver: anti-albumin ab135575 (https://www.abcam.com/albumin-antibody-c-terminal-ab135575.html)
2. Brain: anti-neuron specific beta III tubulin ab18207 (https://www.abcam.com/neuron-specific-beta-iii-tubulin-antibody-neuronal-marker-ab18207.html)
3. Kidney: anti-SLC14A2 ab95365 (https://www.abcam.com/slc14a2-antibody-ab95365.html)
4. Testis: anti-histone H2B ab23913 (https://www.abcam.com/histone-h2b-testis-specific-antibody-ab23913.html)
5. Ovary: anti-ENPP2 (also kidney, placenta, small intestine): ab77104 (https://www.abcam.com/enpp2-antibody-5h3-ab77104.html)

The advantage of this combination of antibodies is that you can use the same anti-rabbit secondary since they were all raised in a rabbit host. Our anti-rabbit secondary antibodies are listed in the link below:
https://www.abcam.com/Search?Keywords=anti-rabbit&fReset=1&pt=-1&source=TopSearch

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

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link:
https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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. This will be the same lot as what you received before. We do not have other lots, but we have also not received other complaints about this one, and it is possible the issue was with the single vial.

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.

Thank you for contacting us about this issue. Your protocol appears to be correct, and you should be seeing a signal with 50ug per lane and a 1/400 dilution. Sometimes switching from milk to BSA for blocking and diluting the primary will help develop the signal more but if you see nothing at all then I am skeptical this will help.

Have you been able to stain blots of these same samples with other antibodies? You mentioned that the secondary was known to be good but it is not clear if you used with other primaries on these HEK 293 lysates, or different lysates. In any case, I think a replacement is in order. Are you interested in trying a polyclonal antibody instead, such as ab82383?

https://www.abcam.com/ENPP2ATX-antibody-ab82383.html

I look forward to your reply.

Thank you for contacting us.
We recommend to run the gel in denaturing and reducing conditions for ab77104. Therefore, SDS PAGE gel should be used. Attached is our WB protocol and you could use it as a reference.
We use HEK 293 (huamn embryonic kidney cell line) as positive control. You could prepare the cell lysate according to the attached protocol or use ab7902 (HEK293 Human embryonic kidney cell line Whole Cell Lysate) as positive control.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for your enquiry.

I am sorry to confirm that these antibodies have not been tested in sandwich ELISA (sELISA). They have been tested in indirect ELISA only and I can recommend considering using them in an indirect ELISA rather than sELISA.

Sandwich ELISA procedures can be difficult to optimise and tested match pair antibodies should be used. Any tested matched pairs would be noted on the datasheet. This ensures the antibodies are detecting different epitopes on the target protein so they do not interfere with the other antibody binding. Therefore, I am sorry we are unable to guarantee our antibodies in sandwich ELISA unless they have been specifically tested for sandwich ELISA.

I am sorry I am unable to suggest any other antibodies suitable for sandwich ELISA for you on this occasion. However, I hope this information is helpful, Should you have any further questions, please do not hesitate to contact us.

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 978396. 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.  

Thank you for taking the time to complete our questionnaire and provide these details. The details you have kindly provided will provide us with vital information for our monitoring of product quality I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been succesful. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably recieved a bad vial. I apologise for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation. Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.    

Thank you for contacting us. Ab82590 and ab77104 are both Anti-ENPP2 mouse monoclonals, however,  ab82590 is from the clone [1F8] and ab77104 is from [5H3]. We are sorry if any confusion has been caused in this matter. We have no plans to discontinue either product. I am sorry to hear you have been experiencing problems with ab82590. This antibody has been tested using mouse and human samples and in paraffin embedded IHC. We would therefore expect your experiments to be working. If you wouldn't mind, could you please fill out the questionnaire I have attached in order to find out a few more details about the protocol used. This may allow me to suggest some things in order to improve the result, and will also allow us to monitor the problem and initiate any internal investigations needed in this case. If the antibody was purchased within 6 months and used according to the datasheet you will be covered by the Abpromise and will be entitled to a free of charge replacement or a credit note. I look forward to receiving your reply.