Anti-alpha Actinin antibody [0.T.02] (ab18061)

Thank you for your email - the problem is that we used same conditions as before (the replaced antibody with long incubation time), however, we did not get any SUR1 bands at all. You think 1:100 dilution is OK to use, but we incubated the blot at this concentration for very long time (3 days), this is quite different from the most other antibodies people used! This time even incubating for 3 days won't work (but beta actin is fine).

We do not know what else we could do?

ANSWER:

We now have new lot in stock so I can send you one vial as a replacement antibody. Again because we haven't tested this lot in mouse so we cannot confirm the ab will work.

You have kindly mentioned in previous emails that the antibody worked before so perhaps running suggested human lysates will give goodindication whether the antibody is wrong or the lysates.

Let me know if you are interested in getting the new lot.

Please see attached - let me know if you need any further info.
thanks,

ANSWER:

Thank you for confirming all the details.

The mouse species has been added to the datasheet from a publication http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=20362548&dopt=Abstract. We unfortunately hold no tested data with mouse lysates. The mouse and human SUR1 are quite similar at the immunogenic region so it is more likely that the antibody will work.

We currently haven't received any complaint other than the 2 you submitted last year. So we are not sure if this issue is lot related. I have checked the case again and would like to suggest changing few things and hoping that the antibody will be OK;

- SUR1 is multipass membrane protein; for mmp we do not recommend boiling the sample with loading buffer. Please try heating samples for 60-70C for 10-15 minutes.
- The lot conc. we have sent is 0.5 mg/ml. 1/100 dilution is OK to use.
- if possible please include a positive control with human cell lines e.g. U2OS, A431, pancreatic tissue lysates, Tonsil, gall bladder, liver or lung tissue lysates.

Please address all the emails of this complaint to me (ab32844).

Let me know how these suggestions go. if the results does not improve I will then ask to choose another anti SUR1 antibody.

Thank you for your email.

Previously we had difficulty with the ab32844 anti-SUR1 antibody (last year), and the replaced antibody was working OK for both WB and ICC (using longer time for incubation in WB, similar to this time, we could detect the expected band). It used up quickly, but that gave us some useful WB data.

We currently need to do more WB in some sets of samples for a manuscript, and hope the newly ordered SUR1 antibody will work (but this lot seems not working).

Please let me know if you need any other information.

Thank you again,

ANSWER:

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

I look forward to receiving your reply.

PS:Please provide all the details of experiemtn andany troubleshooting you have already done! Thanks. Please do not forget to add the purchase order number.

I would like to ask you for help. We need a specific antibody only labeling cardiomyocytes. If possible, the antibody should be labeled already. I have already bought the TroponinI antibody (ab97711) of Abcam which is working quite well. If you could offer another one as desribed, I would be glad. Thank you in advance for your help! Kind regards

ANSWER:

Thank you for contacting us.

There are many proteins that act as markers for Cardiomyocytes; antibodies against these targets can be used for ICC/IF or other required applications.

Main cardiac proteins are as follows;

- alpha actin; alpha sarcomeric Actin antibody [Alpha Sr-1] (ab28052) - N-Cadherin: N Cadherin antibody (ab12221) - alpha actinin; alpha Actinin antibody [0.T.02] (ab18061) - bFGF: FGF basic antibody (ab16828)

I would recommend typing the target name in the search box provided on the Abcam homepage and then selecting the antibodies as per your requirements e.g. monoclonal, polyclonal, rabbit or mouse etc.

The full list of Cardiomyocytes markers can be found by clicking the below link;

http://www.antibodybeyond.com/reviews/cell-markers/cardiomyocyte-marker.htm

Please click on the following link to explore the Abcam Cardiovascular page

http://www.abcam.com/index.html?pageconfig=resource&rid=10694

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

I have a question about your alpha actinin antibody [0.T.02] (ab18061). Does this antibody regognize Rat alpha actinin 1 (SwissProt: Q9Z1P2)?

ANSWER:

Thank you for your enquiry. The immunogen for ab18061 is full-length human alpha-actinin 1 from platelets. There is no epitope information for this monoclonal antibody; however, the sequence homology between human, mouse and rat alpha-actinin 1 is 99% for all combinations.

I hope this information helps you, and please do not hesitate to contact us if you need any more advice or information.