Anti-IP3 receptor 抗体 (ab5804)

I would recommend to follow the protocol we have on our website. The use of 2 times the concentration of peptide (compared to the antibody concentration) represents a large and sufficient excess to block the antibody. I hope this is helpful. Do not hesitate ton contact me again if you have questions.  

Thank you for confirming these details and for your cooperation. As agreed, 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.  

Thank you for confirming these details and for your cooperation. I can confirm that : - the blocking peptide for ab5804 is ab5839 - the immunogen of ab103278 is aa 1155-1184 of the isoform A (SwissProt: Q9NQ66-1) Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products. I am happy to issue a free of charge unit of each the above products. In order to prevent any shipping error, could you please send me the original order number(s), the date(s) when the order(s) was(were) placed or at least the shipping address? Thank you.

Thank you for contacting us and for sending images of the results obtained with ab5804 and ab77743. 1)  ab5804 : I looked at the image of the blot and I am positive that the band observed above the 250kDa marker is ITPR1. As explained in my previous email, Western blotting big proteins of more than 300 kDa is not that easy. From your blot, I can see that the transfer went well. Regarding the size, it looks close enough to 300kDa to confirm you targeted the right protein. The values for the expected as well as the observed MW stated on the product datasheet were a typo and the datasheet has been updated. The expected MW for the Human ITPR1 isoform 1 is 314 kDa (http://www.uniprot.org/uniprot/Q14643). The observed molecular weight should fluctuate around this value but should not be, I think, as low as 240 kDa. In general, the apparent molecular weight is affected by the pH/polarity of the buffer giving a different apparent molecular weight in different buffers. The bands observed around 100 kDa may be due to degradation of ITPR1, if so make sure you add protease inhibitors in the sample buffer. It also may be due to some non-specific binding of the antibody to other protein presenting a sequence similar to the immunogen peptide. It may also be due to some non-specific binding of the secondary antibody, to determine this last possibility it is recommended to run a "no primary" control. Finally, I would suggest to try a different blocking agent. 5% BSA can sometimes reduce the background as well as enhance the specific binding. 2) ab77743 : the recommended dilution is 1/200, have you tried the antibody in this condition? An incubation overnight at 4°C is common. You may also try at room temperature for 3 hours. I would recommend not to use milk as blocking agent for phospho-proteins. 5% BSA is recommended.   I hope these suggestions are helpful. If not, do not hesitate to contact us again and to visit our protocols page at  www.abcam.com/protocols and our protocol book available at : https://www.abcam.com/index.html?pageconfig=resource&rid=13111  

Thank you very much for your inquiry and your interest in our antibodies. Indeed, we do not have ITPR1 antibodies tested against bovine in our catalogue currently. We are however able to provide you with a testing offer, please see below. In regards to the alignments, I do not have the immunogen sequence of the ab108517 as it is proprietary information of the laboratory from whom we source this antibody. I have however requested the alignment information from them. Please see here below also the alignment of the immunogen sequence of ab97823 with the bovine sequence. The alignment is very good with 93% identity. The alignment of the immunogen of the ab5804 has however 100% identity with the bovine protein, so crossreaction is even more likely. In regards to the blast, I am sorry that I do not really understand the question. I have blasted the immunogen against the human proteins (to have a better overview) and have found only significant reaction with the ITPR1 protein. There are different isoforms, however all from the ITPR1. Please provide me maybe with the relevant sequences. We can provide you also with a testing offer for testing an ITPR1 antibody against bovine. Indeed, even if the immunogen sequence is identical, we do need experimental evidence to be able to guarantee it. Therefore as to our knowledge, ab97823 or ab5804 have not been tested in bovine, I can offer a discount off a future purchase if you buy ab97823 or ab5804 now, test it in bovine and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody. This offer might also be applicable to the ab108517, we need however to wait for the alignment results in order to be able to decide on this. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and test ab97823 or ab5804 in bovine. I will then send a discount code. This code must be issued before purchasing ab97823 or ab5804 so please wait for my reply before ordering. 2. Purchase ab97823 or ab5804 either by phone, fax, or online (www.abcam.com). 3. Test it in bovine. 4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out more about our Abreview system, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab97823 or ab5804 turns out to be unsuitable for bovine, you will still receive the discount on your next purchase after your Abreview has been submitted. Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.