I am currently in the process of doing a Western Blot with mESCs after an AHA Assay. I added 25uL of 2xLaemmli buffer and 25uL of AHA buffer (50mM Tris HCL) to my pellet.

I usually denature my proteins before loading on SDS Gel by boiling the sample with dye at 95C for 5 min. However, this time I forgot to take only half of my sample and store the other half for a repetition in the future. Instead, I added the dye and denatured the protein lysates by boiling. Can I still load half of my sample and save the rest in -80 or -20? Or should I leave it at RT? Or finally, should I just discard the rest of my samples?

I will appreciate some help!