Anti-Lin28 antibody (ab46020)

Not seeing the expected band size in cells containing constructs of human Lin28A or Lin28B. Does this antibody target Lin28A or B?

ANSWER:

Thank you again for your call yesterday and for your patience.



I've heard back from the lab about ab46020, and this antibody was raised against full-length Lin28A. It has not been tested specifically against Lin28B but since these proteins are so similar, it is expected to cross-react.



In light of this information, I'm still not sure how to interpret the data with this antibody. I'm hoping that the new antibody will provide some informative data. Please keep me updated about the results with this antibody.



I look forward to hearing from you. Please let me know if you have any questions or if there is anything else that we can do for you.

Not seeing the expected band size in cells containing constructs of human Lin28A or Lin28B. Does this antibody target Lin28A or B?

ANSWER:

Thank you for your call today and for letting us know about the trouble with ab46020. I apologize for the problems with this antibody and for the inconvenience.

As we discussed, I'm sending a free of charge vial of ab71415, which is specific for Lin28B but is predicted to cross-react with Lin28A. This vial should arrivetomorrow on the order ***. Please keep me updated about the results with this replacement antibody.

I'm also getting in touch with the lab to request more information about the specificity of ab46020, and I will forward that information to you as soon as it's available.

Please let me know if you have any questions or if there is anything else that we can do for you, and I'll be happy to help.

I have searched the Abcam online cataogue and found specific antibodies for lin-28 (ab46020) I would like to know if it is specific for the lin28A or lin28B. Also, there's the specific antibody to detect lin28A in the antibody list, but are there any specific antibodies which probe for lin28B? Thank you for your considerations and help if you may. I look forward to hearing from you soon.

ANSWER:

Thank you for contacting us. Ab46020 was raised against full length Isoform A. We predicted that it could cross-react with isoform B, however, we have not tested it in the laboratory. In our catalogue, there are totally 3 items which can specifically react with with human LIN28B, they are: ab71415, ab115698, ab119367. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Hello, the problem is, that there was no staning by use manual staning method - three different permeabilization steps (citrate buffer, high pH, EDTA). By use automatic staining system Benchmark XT, we got positive signal with very strong dilution 1:50 and 1:75 - but with incorrect nuclear localization a background staining. The staining with dilution 1:100 and 1:150 wasn´t so specific, how we expected. Please, see the attached pictures. with best regards  

ANSWER:

Thank you for taking the time to complete our questionnaire and provide these details.

The details you have kindly provided will provide us with vital information for our monitoring of product quality I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been succesful.

Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can suggest you have regrettably recieved a bad vial. I apologise for the inconvenience and am pleased to offer you a free of charge replacement or credit note in compensation.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.