Anti-Cryptosporidium抗体[BEL 0126] (ab54066)

Thank you for your reply.

The volume added to the wells depends on the number of cells and surface available. It should cover all the cells surface completely.

You may want to try less concentrated antibody dilutions, for that, I would suggest trying first with a small amount of sample and antibody to verify the specific dilution is enough to provide a visible signal.

I hope this helps, otherwise, please do not hesitate to contact me back.

Thank you for contacting us.

I appreciate the time taken to submit protocol information. The details you have kindly provided will provide us with vital information for our monitoring of product quality. I would also appreciate if you could please send us an image and the order details.

Reviewing this case, I would like to offer some suggestions to help optimise the results from ab54066:

Fixing in paraformaldehyde for more than 10-15 min will cross link the proteins to the point where antigen retrieval may be required to ensure the antibody has free access to bind and detect the protein.



Instead, I would suggest fixation using 4% paraformaldehye to slides for 10 min only, or even better, trying acetone fixation, at -20C for 5-10 minutes. As this agent will also permeabilise, further permeabilization steps will not be required.

Use higher antibody dilutions (1/50) and use a positive control to assess the antibody’s performance.



I would suggest having a look at the following protocol, which will hopefully serve as a guidance to optimise the assay: https://www.abcam.com/index.html?pageconfig=resource&rid=11417

I hope this information is helpful. Should the suggestions not improve the results, please do not hesitate to contact me again and I will try to provide further help.