Question (28891) | Anti-Doublecortin antibody (ab18723)

Go to datasheet (ab18723)

Question

We have recently purchased one of your anti-doublecortin antibodies (Ref ab18723, lot numberXXXX) in order to characterize neurogenesis . We use rats, perfuse them with PFA 4% for 7 minutes and postfix with the same fixative for 3h, after two washes in PB, samples are cryopreserved in 30% sucrose and frozen with isopentane. Sections are 40 microns thick and stored free floating in antifreeze solution, IHC is performed in freefloating. We used 1:1000 dilution in blocking buffer containing 10% NCS over night at 4ºC and detected signal with secondary antibody coupled to DyLight 594. The obtained signal is quite strange to us, as many cells look like astrocytes (see picture attached), do you have any information or suggestions that could be useful to us? We processed samples without primary antibody so we are sure the secondary Ab it is not the problem.

Answer

Thank you for contacting us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.  Regarding the image sent it seems to be high background and no specific staining on the samples. In order to improve the results from this antibody, I would like to offer a suggestion. It would be worth using different more diluted antibody dilutions, to try reducing the background staining. I would suggest 1/1500 or 1/2000.  I would also appreciate if you could send me the order number, so I can check there has not been any issue during the shipping. In any case, if the antibody was purchased within the last 6 months, it is covered by our Abpromise guarantee, and therefore, I’ll be more than happy to offer you a free replacement of ab18723 form a different lot in case there has been any problem with this particular lot. I hope this information is useful. Please do not hesitate to contact us if you need further advice or information.

Sign up