Anti-Cytokeratin 19 抗体 (ab15463)

Thank you for taking time to contact us. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

This antibody hasn't yet been validated in pig, so we are unsure whether or not it will work in this species. Would you have access to run a positive control with mouse or human skin?

Is the trouble that you are not seeing any staining? If you aren't seeing any signal, you may want to cut down on the blocking percentage and time of incubation.

We have heard that epitope retrieval with citrate buffer ph 6.0 for 10 minutes at the microwave at 600 W and blocking the endogen peroxidase with 3% H2O2 in Methanol for 10 min, with primary at 1/50 overnight 4C works well. The secondary was used at 1/200. What dilution of the secondary have you tried?

Should the suggestions not improve the results, please do let me know.

I hope this information is helpful, and I thank you for your cooperation.

Thank you for taking the time to complete our questionnaire and sending me the publication. I am sorry to hear you have had difficulty obtaining satisfactory results from these antibodies.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.


Although the other antibody has worked well using this procedure, individual antibodies will often require optimization. I would like to offer some suggestions to help optimise the results from ab15463 and ab37994.

1.) I suggest to use a different positive control than HUH7 cells. As shown in figure 1 of the publication provided HUH7 cells do not show any expression of CK-19 and a only a very low expression of DCAMKL-1. I can recommend to use HepG2 cells or shin for ab15463 and MCF for ab37994 as indicated on the respective datasheets.

2.) I also can recommend to incubate primary antibodies over night at 4C. This will ensure the best saturation and specificity of the antibody.

3.) For the WB, I suggest to use less than 10% milk powder as blocking agent (maybe try a different blocking agent like BSA which can sometimes have a big impact on signal strength).

4.) For ICC/IF I can recommend to try a milder permeabilisation agent./ For example 0.1 Tween in the blocking buffer for 1 hour.


I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again.