Hello,
I have a very odd problem. A few years back (2016) I was using MyOne Streptavidin C1 dynabeads and coupling them to a biotinylated peptide and doing pulldowns using whole cell lysates. My pulldowns were working really well and the beads alone control never showed any background binding by WB or Coomassie staining.
Fast-forward 3 years and the beads no longer perform well for me. The same exact experiment using the same exact protocol (buffers and biotinylated peptides) has the same amount of enrichment on my beads alone control as all my test samples suggesting that the beads now bind to everything unspecifically. I have contacted customer services about this and they have sent new batched of the beads yet the problem persists. It is ridiculous as I can no longer perform my experiments and I cannot explain why the beads have suddenly become so sticky. I tried changing salt concentration to see whether I could change the background binding but I had no luck. These beads have become totally useless. However, M280 streptavidin and MyOne Streptavidin T1 beads do not show this unspecific background so I am confident the problem is not my protocol. Unfortunately, these beads also don't perform as well as the original beads in the actual pulldown samples either so i am reluctant to move over to these on top of the fact that I have a lot of solid data obtained from the MyOne Strep. C1 beads that I do not feel comfortable switching beads half-way through a project.
I was wondering whether anyone is still using these beads successfully.
Thanks in advance :)
Julia
Hi Julia,
sorry to hear about your challenges. The properties of the beads should not change like this.reach out to me at [email protected] to see what we can do.
kind regards
ketil
Dear Ketil,
Thank you for your response. I have already been in extensive contact with someone from the Thermofisher customer service and we are still trying to solve the problem.
I was just wondering whether other researchers using these beads had had a similar experience to me.
Thank you for your offer of help.
Regards,
Julia
I am facing problem with Dynabead M270 carboxylic acid. I have followed the exact recommended protocol and recommend (and twice) concentration of peptide (38AA). For incubation I performed room temperature and overnight at 4 degree both. But my peptide was not immobilized onto bead. I got same concentration of peptide in supernatant which i used initially for ligand immobilization. Any suggestion is highly appreciated.
Did anyone get an answer to this? I have found a similar issue, whereby I am struggling to get bid efficient binding of my biotinylated proteins to MyOne Streptavidin C1 dynabeads. This is a protocol that has been used many times in the past and analysis of my output sample shows that most biotinylated proteins are not pulled out by the streptavidin.
Hi Julia, Julia Mawer
I am currently using the same beads Dynabead myOne C1. I have tissue samples that are limited and have tried them on other more availbale tissue first (for pul-down of endogenously biotinylated proteins). MS analysis have showed that they indeed pull-down biotinylated proteins. However, I came across your question and I was wondering whether you have solved the problem or got to know what caused them to be so sticky?
Any answer would be highly appreciated.
Regrads,
Zeynep
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