any suggestions ragarding tricine gels - (Dec/01/2008 )

hi
my protein/peptide of intrest looking for resolving is 2 kd
i have been trying in tricine sds page
but below 4kd nothing was there in gel yet..,not able to resolve below 3-4 kd
plerase suggest me to resolve my 2 kd peptide
any additions or deletions or modifications are required
if so please suggest me the possiblities
thank you

what percentage gel did you use?

for 2kDa i would use at least 16.5% acrylamide (maybe 20%) or a 10-20% gradient.

also

staining may be difficult (depending on constituent amino acids). ensure you use a method that can detect your peptide.

for a gradient you need to prepare gel solutions at the starting and ending concentrations (approximately 1/2 of the gel volume for each concentration, approximately because you can, and should, make volume adjustments based on density differences and starting pour).

using a gradient former, and pouring from the top, start pouring the denser (higher concentration) solution into the sandwich. when the solution reaches all the way across the bottom, open the valve between chambers, allowing the lighter solution to enter and mix with the denser solution (do this with mixing).

when it is finished pouring, overlay and allow to polymerize then rinse, stack, and run.

if you pour from the bottom up then start the lighter solution first.

add aps just before you pour.

or you can buy gradient gels.