Anti-M6PR (cation independent)抗体[MEM-238] (ab8093)

I have issued a free of charge replacement with the order number for ab32815.

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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, I received the data this time. For a protocol modification, I suggest trying preparing a sample by heating at 60C for 10 minutes instead of 95 for 5 minutes to minimize aggregation of the trans-membrane domains. It is strange that both antibodies give you this result, and I still think there may be some issue with the sample entering the gel and/or transferring effectively, even though the standards do.

Another modification is to remove methanol from the transfer buffer and add SDS to a final concentration of 0.1%, to encourage transfer of this large protein.

Are you confident that the secondary antibody is not reacting with the samples? Is the secondary specific for a subclass of mouse IgG?

These modifications are unlikley to remove the band at 70 kDa. We do have other antibodies against the receptor, so if these modifications fail, I can send a vial of something else to try, such as ab32815 or ab124767.

Thank you for bringing this to our attention. I will need a few more details of your protocol. I was not able to open the file you sent.

Can you please tell me how you prepared your lysates (did you use protease inhibitors), and how you heated the samples to denature. I am concerned that the receptor may have aggregated upon boiling and did not enter the gel effectively, or did not transfer to the blot well. Did you check for efficacy of transfer with a Ponceau Red stain or equivalent?

How much sample did you load per lane of the gel? How did you block the membrane? What concentration of antibody did you apply to the membrane and how long did you incubate with the antibody?

I'm glad that you have been seeing better results with blocking with BSA. If you are now having difficulty with non-specific bands, increasing the dilution of the primary and secondary antibody as well as adding 1 % BSA and 0.2% Tween 20 into the diluents may help. If you would like me to have a look at your protocol to see if there is any other advice I can think of then please do fill in the form and I'd be happy to try and help. Otherwise, I wish you all the best with your experiments.

Thank you for contacting us. Having looked into the case a little I can see no reason why ab8093 should not be performing well with Western blotting or that ab2733 would be likely to perform any better. What might be helpful is if you were to share the protocol which you are currently performing and I may be able to spot some things which may help to improve the performance of ab8093. To this end I have attached a questionnaire, this should only take 5-10 minutes to fill in. It would be helpful if you could also attach an image of the blot obtained. This information will also help us to monitor the quality of our products and to determine if any further internal testing needs to be performed. If the antibody is not performing to the level we would expect with the applications and species listed on the datasheet you would be eligible for a replacement antibody or refund. We do have an antibody in our catalogue which have been more fully characterised with Western blotting and may serve as an alternative to ab8093 if necessary. Ab32815 has been used by one of our customers to detect the Mannose 6 Phosphate Receptor protein from the membrane fraction of Hela Cells using Western blotting: https://www.abcam.com/M6PR-cation-independent-antibody-ab32815.html&tab=abreviews&intabreviewid=11713 I'm sorry that I could not be of more help but hopefully with the protocol in hand we can decide if trying an alternative antibody would be worthwhile. In the meantime I am sorry for any inconvenience this has caused.

I have attached an Excel file with information regarding all our current antibodies that have been tested in EM. I hope this is helpful. Please let me know if you have any further questions.

Thank you for your enquiry. I am working up an internal query to obtain a list of all EM-tested antibodies in our catalog. I will forward you this list as soon as possible. Thanks for your patience!