Samples can be shipped to:

Whitehead Institute
455 Main St
Rm327/ Proteomics Facility
Cambridge, MA 02142
617-324-1805
Eric Spooner

• !For overnight shipping it would be great if samples could be shipped so that they arrive on Tuesday or Wednesday to ensure they are stored properly upon arrival. Priority overnight will ensure the samples arrive late morning which is greatly preferred and will ensure your samples are handled by Proteomics Facility Staff upon arrival.

• For sammples that can be hand delivered to the building please make arrnagements with Ed Dudek or Eric Spooner and we can arrange to meet you at the building lobby

Gel-based Samples

• Coomassie Staining
  
  • Coomassie stained gels can be destained, washed and submitted as below. Both standard coomassie stains and coloidal coomassie stains work well and have more than enough protein for good sequence coverage. Both commercially available kits and homemade solutions are fine. Coomassie stained gels are preferred for post-translational modification determination.
  
  
• Silver Staining
  
  • !!It is critical to not overstain gels in order to obtain sufficient peptide recovery.!!
  • Silver stain is a covalent stain which attaches to the polyamide backbone of proteins. The staining should be on the face/surfaces of the gels.
  • Peptide recovery comes from the unstained acrylamide in the middle of the gel.
  • Sigma or Invitrogen silver stain kits work fine as long as they are not real old. Avoid BioRad kits. You should skip the destaining step.
  • We recomend the following Silver Stain Protocl which is commonly used at the Whitehead with great success. (The initial cost for the reagents is ~$200 and you can stain many gels if you choose to use this in the future).
  • This is extra information detailing some of the finer points of the protocol.

Once you have stained the bands, you can excise them, cut them into smaller pieces (~2-4 mm squares), place in a 1.5 ml Eppendorf and cover with 50/50 Methanol/Water. These can be shipped overnight to the above address (dry ice is not necessary.) Feel free to give me a call or follow up email if you have any other questions.

Solution Based Samples

You must discuss with Eric prior to submitting samples if you have not previously submitted solution based samples!

• Samples need to be at a pH of 7.5 - 8.0 for tryptic digestion. Some salts and detergents can be tolerated.
• TCA precipitation and resuspension in a Tris/ Urea buffer can be used for samples not amenable to tryptic digestion.
• Samples should be shipped overnight with dry ice/cold packs.