We are working with stably integrated AR in Hepg2 cell line. While doing characteristaion we are not able to get band for our desired protien of 110 kda in WB. However, other confirmatory results of ICC, PCR and transcription assays show positive data of GOI to be integrated in cell line. I ahd tried the ab dilutions from 1:2000 to 1:10000. Every time we get a single faded band at 97 kda. I do blocking with 5% milk in 1X TBS. AB for GOI is polyclonal. I perform transfer at 25 volt for 2 hours and had tried to extend the transfer time to 3 hours but with no gain. I will be uploading my blot images for review. Any leads would be very helpful. Thanks in advance.
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