Anti-R Cadherin 抗体 [EPR4836] (ab109242)

Thank you for the protocol and sample details.

The 100kDa band may be cadherin. The size of the protein, without any post-translational modification such as glycosylation, is expected to be 100 kDa, according to the UniProt database entry for the mouse CADH4. Are these osteosarcoma cell lines known to glycosylate CADH4? Here is a link to the entry. The amino acid sequence and size are listed far down the page:

http://www.uniprot.org/uniprot/P39038

The 80 kDa band may represent a fragment. Can you please confirm that you collected your samples with protease inhibitors? Have you tried staining blots of these samples with any other antibodies?

The recommended dilution is a range from 1/10,000 to 1/50,000. The antibody has very high affinity for R cadherin, so I recommend stripping a blot and re-staining with a more dilute solution of the antibody, for instance 1/25,000. This should reduce the amount of background staining.

Please let me know if you have any questions or concerns.

Thank you for bringing this to our attention. Can you please provide some details of your samples and protocol? For instance, what is the species and tissue type or cell type, how were the samples prepared, how much sample did you load per lane, what did you use for a blocking buffer, and what was the dilution or concentration of the antibody? Are you sure the secondary antibody is not the problem?

Can you please also send the order details? The name of the institution and the approximate date of the order will be sufficient.