Anti-SIRT2 抗体 (ab19388)

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, the rabbit monoclonal ab134171.

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 your reply, the refund will be arranged soon by our accounts department. I'm afraid we do not have the immunogen peptide in our catalogue nor can we source it for you as ab19388 is not made in house, I'm very sorry I cannot help you more,

I'm sorry to hear you are experiencing problems with ab19388. Many thanks for providing the image of your blot and your detailed protocol, it is very useful for me to understand your problem. I would like to recommend some changes to your protocol to increase the signal specific to SIRT2 and to decrease the background too. I am not sure how high levels of SIRT2 are in the adult brain so I would recommend to run a positive control along your samples, such as mouse 3T3 lysate. Is the species you are using human or rat? I am also concerned that your samples date from last October and were stored at -20C if they were stored in lysis buffer. If they were stored after boiling in loading buffer they should be fine. Were there plenty of fresh protease inhibitors at the time of lysis, could they have gone off? I would recommend to incubate the membrane at 4C overnight and to try a range of dilutions (you may be able to dilute more by incubating longer), this will give a slow but targeted binding. To improve on the background on the membrane I would recommend to try blocking 1hr only in BSA (5% in TBST) and to incubate the antibodies in TBST only, at 4C. We recommend to use HRP conjugated secondary antibodies and ECl+ detection systems, so if you have those available it may be worth trying too, as they give less background. Please also wash the membrane in TBST only, this should help too. I hope the above recommendations will help you, please do not hesitate to contact me again if you need further assistance, ***An error on the datasheet regarding testing in mouse samples has since been detected and the customer was refunded her order. Mouse samples have not been tested***

Thank you for contacting us for technical support. The fact the customer is having the correct results with over-expressed protein reassures us that the antibody works and that in Hek293T cell lysate the protein levels are much lower. We have not had any complaints about these antibodies and they work very well in the tested positive controls. I would recommend running the positive controls detailed on the datasheets: ab10140: human kidney cell lysate, U937 lysate ab19388: 3T3 cell lysate ab13860: no cell line has been tested- please try human testis lysate ab13697: no cell line has been tested- please try human intestine lysate If you require further assistance can you please ask your customer to fill in our protocol questionnaires and we can look at each antibody condition; typically please check your customer incubates at the recommended dilution in TBST overnight (he/she may need to use more concentrated antibody as this depends on the tissue), please check that degradation of the protein is not occurring and other protocol details which may make a difference,

Thank you for your email, and I'm replying on behalf of Sarah who is currently away from the office. I'm sorry to hear that ab19388 is still giving you difficulty and I can certainly arrange for a free of charge replacement vial to be sent to you. As you ordered via one of distributors, Biozol, please notify them of the situation (forward my email to them) and they will send you the replacement. Please let us know how the replacement works out for you and if you have any additional questions.

Thank you for contacting us for technical support and for taking the time to provide all your protocol details, this was really useful to understand your problem. I suspect the problem is due to the blocking agent present in your antibody dilution buffer. I would recommend trying: -5%BSA 1hr then incubating the primary and secondary in TBST only -5% milk 1hr then incubating the primary and secondary in TBST only This will give sufficient blocking to prevent non specific binding but we know that too much blocking can also cause non specific binding, so a balance is essential. The rest of your protocol looks really good so I think you will have much better results with this change (though you can also try to load a little less sample and dilute more your secondary and do more washing steps too) (also make sure the secondary works well with other primary antibodies). Please let me know if you still experience a problem and I can offer you a replacement vial or refund if you purchased the antibody in the last 90days