100 ul is quite a lot, If you are experienced gel runner even 30 ul is quite a lot, generally to get sharp crisp bands the lower the loading volume the better, you may make an argument about the stacker, still lower volumes are good. after boiling spin the tubes, so all settle well down at the bottom. Again if experienced go for 20 ul, if not 30 is good. after boiling you would lose some in evaporation anywhere from 5-10 ul. Good Luck with results.

Mary

100 ul is quite a lot, If you are experienced gel runner even 30 ul is quite a lot, generally to get sharp crisp bands the lower the loading volume the better, you may make an argument about the stacker, still lower volumes are good. after boiling spin the tubes, so all settle well down at the bottom. Again if experienced go for 20 ul, if not 30 is good. after boiling you would lose some in evaporation anywhere from 5-10 ul. Good Luck with results.

Mary

30uL Laemmli buffer is better option, however I would suggest to load only half of it to a single well.

As said above, 30-40µl is fair enough! If you experience multiple bands while developing blot, then dont follow this method of heating the beads directly, instead do elution by competing molecule(Imidazole in case of NiNTA beads) and use that eluent as your sample. Personally I tackled this issue by doing this.

Good luck.

100ul of _beads_ is a lot. Do you need that much- considering the high specificity of streptavidin? If you are only expecting to obtain enough target protein fro one lane of a gel then you probably don't need to use so many beads. just a thought

As mentioned by Desmond above, If it is not a preparative purification, 30-40µL of bead slurry should be enough for Strep PD. then u can easily elute it using 30-40µL loading dye and load in gel. It will be tough to get some liquid to load, if u add 30µl in 100µl bead slurry after PD