maldi compatible silver staining problems

[bsengez]

Hi everyone,
I have silver stained 2D-gels, but unfortunately I cannot get a good MS/MS result. Is there anyone using silver staining and getting results in MS/MS? Thank you in advance..
Burcu

[admin]

Hi there,
usually people don't use silver staining and then do mass spec unless its a compatible silver stain, and your mass spec facility can do it.

There is a Pierce silver stain kit compatible for mass spec, but if you already stained it - let the mass spec facility know your steps - they should be able to troubleshoot and help.

Let us know what happens next,

[bsengez]

I also tried to use coomassie even colloidal coomassie staining, but unfortunately I have poor results. Therefore, I prefer to use silver staining. I'm also carrying out mass spec analysis. My problem is there are several mass compatible silver staining protocols and I need the best one. For instance, some protocols use formalin in each step but the others use it only in developing step. Some uses only EDTA in stopping step, the others acetic acid. Therefore, I need people who have good mass spec analysis using silver stained spots.
Regards..

[Warthaug]

You can silver-stain, but you must destain the cut-out bands or the mass-spec will not work. Your mass-spec facility may have the methods to do this - its quite a long procedure and I don't have a protocol on-hand.

Bryan

[bsengez]

yeah I know, before in gel trypsin digestion I am also destaining it, but there is a still problem.

[arnab_roy]

Hi bsengez, We have done silver staining of SDS-PAGEs of BSA and got well enough MS result as well as MS/MS. But as the concentration of the protein comes down, getting MS/MS done in MALDI becomes tougher. Please post your detailed protocol. May be it will help.

[Mint]

my lab uses silver-stained gel plugs for protein ID, so far no problem, 100% successful rate. But you have to use MS compatible silver staining method, where no glutarldehyde and formaldehyde in silver staining process.

[biologists]

Online consult on bioproximity[dot]com