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Frequently asked questions
Protein, detection & labeling
Protein, detection & labeling
What is the recomended buffer composition for protein electrophoresis using iD PAGE gels ?
iD Buffer composition
5x Sample Buffer
SDS-PAGE | Native PAGE | Final concentration (1x Buffer) | |
Tris base | 300 mg | 300 mg | 50 mM |
---|---|---|---|
Glycerol | 5.0 mL | 5.0 mL | 10% |
Bromophenol blue | 25 mg | 25 mg | 0.01 % |
2-mercaptoethanol | 1.0 mL | 1.0 mL | 2% |
SDS | 1.0 g | - | 2% |
Deinonized water | to 10 mL | to 10 mL | - |
pH (use 8M NaOH or 8M HCl) | 6.8 | 6.8 | - |
Store at room temperature
1x protein sample solution:
Sample | x µL |
Sample Buffer 5x | 2 µL |
Deinonized water | to 10 µL |
Heat at 95°C for 10 minutes before loading on gel
10x MOPS Running Buffer: to separate medium to large proteins >20 kDa
SDS-PAGE | Native PAGE | Final concentration (1x Buffer) | |
Tris base | 60.6 g | 60.6 g | 50 mM |
---|---|---|---|
MOPS | 104.6 g | 104.6 g | 50 mM |
EDTA | 3.0 g | 3.0 g | 1 mM |
SDS | 10.0 g | - | 0.1 % |
Deinonized water | to 1000 mL | to 1000 mL | - |
pH (do not adjust) | 7.7 | 7.7 | - |
Store at 4°C up to 6 months
10x MES Running Buffer: to separate small proteins (2-50 kDa)
SDS-PAGE | Native PAGE | Final concentration (1x Buffer) | |
Tris base | 60.6 g | 60.6 g | 50 mM |
---|---|---|---|
MES | 97.6 g | 97.6 g | 50 mM |
EDTA | 3.0 g | 3.0 g | 1 mM |
SDS | 10.0 g | - | 0.1 % |
Deinonized water | to 1000 mL | to 1000 mL | - |
pH (adjust with 8M NaOH or 8M HCl) | 7.3 | 7.3 | - |
Store at 4°C up to 6 months
20x Transfer Buffer:
20x Transfer Buffer* | |
Tris base | 60.6 g |
---|---|
Bicine | 81.6 g |
Deionized water | 1000 mL |
To make 1L of 1x transfer buffer: mix 50 ml of 20x transfer buffer, 100mL of methanol or ethanol and 850 mL of deionized water.