Can I please get any assistance with the optimization of Nrf2 Western Blot in Zebrafish homogenate?

12 December 2018 0 10K Report

I am running a western blot on a zebrafish homogenates and trying to measure the Nrf2 protein and I am lost with the technicality of the method. Only two concise references out there and the troubleshooting is not going so well. Do you have any idea what is the optimal voltages for the running and transfer and the times, the primary and secondary Ab dilutions, does it matter if I use ALP-anti rabbit or anti-mouse (B-actin), and does it matter if I use if I use ECL or BCIP/NBT? Can you please if help me out if you're able to. The bands are just not developing.

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