Anti-muscarinic Acetylcholine Receptor 1 antibody (ab3480)

I am very glad receiving your letter .I have two question,I have bought two kinds of antibody from abCAM, the one is Rabbit polyclonal to muscarinic Acetycholine Receptor(ab3408),the other is Mouse monoclonal to Angiotensin II Type1(ab9391).

Question1:For antibody ab3480 ,there are too many non-specific bands.In your product datasheet ,it tells me that the band is about 64KD ,but I can only detect a light band in that position .I will show you the picture in my E-mail .Because there dosen't have a control . When I overexpressed pCDNA M1 plasmid ,the band at 64KD is a little heavy , but the result can't be repeated every time.The protocol is according to the datasheet .

Question2:For antibody ab9391 ,the question is the same with ab3480 ,Ican only detect a light band at 45KD ,when I overexpressed pCDNA AT1 plasmid in HEK293 cell , the band is also very light .

I hope you could answer me as soon as possible

ANSWER:

Thank you for contacting us regarding ab3480 and ab9391. I'm sorry to hear both antibodies are currently not working well for you.

The problems could be due to a number of reasons and without details of your protocol it is very difficult for me to diagnose and make suggestions to improve your signal. The problem could be due to the way you overexpress the protein and therefore it would be useful to run a positive control lane for each protein. Please note that ab3480 has only been tested in mouse, while ab9391 has been tested in human and rat; if your protein is not from a species tested the problem could be species related and again a positive control will help you determine that.

If you would like us to look at your protocol in details please do not hesitate to contact us using the form accessible below ; this will take you to a short questionnaire to help you put your protocol information together,

http://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=3480&mode=questionaire

thanks for the e-mail. Can you tell me the conditions used for the demonstration western blot for ab3480? i.e. Was it using recombinant protein (how much), or a cell extracted muscarinic type 1 receptor.

ANSWER:

Thank you for your enquiry. To use this antibody in Western blotting, we recommend using it at a concentration of 2 µg/ml and referring to our general Western blotting protocol (located in the protocol section of the online datasheet). To our knowledge, customized protocols are not required for this antibody. If you have any more questions, please do let me know.

I am writing to enquire about your muscarinic acetylcholine receptor 1 antibody (ab3480). We have been looking for such an antibody to do western blotting, ICC, and IP against rat and human forms of the receptor. However, we have been unable to obtain an effective antibody.

Your web-site declares that a refund will be offered if the antibody does not perform. Is this antibody therefore garanteed for western blots against rat and/or human forms of the receptor?

ANSWER:

Thank you for your enquiry. All the information we have on species cross reactivity is specified on the datasheet, these are updated as soon as any new information is brought to our attention. Ab3480 is expected to cross-react with Human and Rat due to sequence homology; however, this has not yet been tested to our knowledge and so we would be unable to provide a guarantee.

Should you decide to go ahead and purchase this product, please let us know how you get on and in return we will forward a reward of your choice, typically an Amazon gift voucher.

Please let me know if you require any further assistance.