Coomassie staining overnight did not work with precast gels?

How much time we need to keep the 12.5% polyacrylamide gel in Coomassie blue stain to see the smaller bands (<5kDa) before overnight destaining? (I know the Coomassie blue staining protocol. I am asking for your practical experience)

Sometimes I prepare a standard SDS-PAGE gel for westerns, wrap with a wet paper towel and cling wrap, and store it in a zip lock baggy in the fridge. Does storage for a few days alter the effectiveness of the gel in terms of sample separation, transfer, or background on the final image?

I know that our proteomics core claims that SDS-PAGE gels used for mass spec should be freshly made. Has anyone ever tested this for westerns?

Hi, colleagues, can I leave a SDS-PAGE overnight after running? Should I leave it in fixation solution (50% methanol, 10% acetic acid) or shall I perform the fixation for 1 h and then leave it in water? Thanks for the help.

by the experiments we have observed that when using conventional destaining method (100ml methanol 100ml glacial acetic acid and 800ml water) initially we get some bands in our gel but when they were kept in water after 48 hours we have seen some extra bands that were not observed initially so what mechanism is there and which destaining technique we should use?

Which is a better option to run SDS PAGE gel...constant current or constant voltage.

Please mention the values and give a reference if possible.

Dear All,

Is it ok to leave membrane incubating with primary antibody (dilution with blocking buffer) at cold room (4°C) for a few day?

I usually incubate primary overnight, but sometime it's on Friday and I prefer continue on next Monday. If I incubate the membrane with primary antibody for weekend, will the membrane has more background or need more wash? even incubating antibody dilluted in blocking buffer (5% milk or bsa)?

Thank you

Cheerios,

I recently read that there should be <5C melting temperature difference between primer pair of PCR. Why?

I have stained my SDS-PAGE with coomassie blue but bands are very light. Probably, the stain was not good. Can the gel be stained again with new coomassie or silver stain?

Will the staining be efficient?

I would like to harvest Photobacterium and Morganella via centrifuge. Which rpm and time is better to avoid damage cells?