Hi all,
I am using the Ni-NTA magnetic agarose bead from Thermo (Pierce™ Ni-NTA Magnetic Agarose Beads) to do the IP, I want to ask in the final elution step, could I just boil the bead with loading buffer for SDS-PAGE instead of elute it? I don't want to reuse the bead and my sample concentration is too low, elution (>200uL) would too dilute.
Should I boil the beads with protein, then use the magnet stand to remove the bead and use the supernatant for SDS-PAGE? will the protein release from the bead like this? Or I could use the whole boiled beads containing protein for SDS-PAGE? Thanks ahead for any answer!
hello,
I believe it may be better to take the beads , add imidazole to them, boil them with the loading buffer and then load the whole mixture on SDS-PAGE.
it is not necessary to boil magnetic beads..but if you are doing it than it will good but proper wash is necessary.
Hi Huiwen,
Have you tried the experiment with boiling the magnetic beads in this way?
I am also running a CoIP experiment with the Pierce Crosslink magnetic IP kit however I have not cross-linked my antibody to the bead. I also would like to avoid the elution step and boil the sample straight away so if you could tell me if you have had success in doing this that would be great.
Thanks,
Alice
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