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Recipes

NuPAGE® MOPS 

SDS Running 

Buffer 

50 mM MOPS 

50 mMTris base 

0.1% SDS 

1 mM EDTA 

pH 7.7 

1.To prepare 500 ml of 20 X NuPAGE® MOPS SDS Running Buffer, dissolve the 

following reagents to 400 ml ultrapure water: 

MOPS104.6 g 

TrisBase60.6 g 

SDS10 g 

EDTA3.0 g 

2.Mix well and adjust the volume to 500 ml withultrapure water.

3.Store at +4°C. The buffer is stable for 6 months when stored at +4°C. 

4.For electrophoresis, dilute this buffer to 1X with water (see page 15). The pH of 

the 1X solution is 7.7. Do not use acid or base to adjust the pH.

NuPAGE® MES 

SDS Running 

Buffer 

50 mM MES 

50 mMTris base 

0.1% SDS 

1 mM EDTA 

pH 7.3 

1.To prepare 500 ml of 20 X NuPAGE® MES SDS Running Buffer, dissolve the 

following reagents to 400 ml ultrapure water: 

MES97.6 g 

TrisBase60.6 g 

SDS10 g 

EDTA3.0 g 

2.Mix well and adjust the volume to 500 ml withultrapure water.

3.Store at +4°C. The buffer is stable for 6 months when stored at +4°C. 

4.For electrophoresis, dilute this buffer to 1X with water (see page 15). The pH of 

the 1X solution is 7.3. Do not use acid or base to adjust the pH.

NuPAGE® Tris Acetate SDS Running Buffer

50 mMTricine

50 mMTris base 

0.1% SDS 

pH 8.24

1.To prepare 500 ml of 20 X NuPAGE®Tris-Acetate SDS Running Buffer, dissolve 

the following reagents to 400 ml ultrapure water: 

Tricine89.5 g 

TrisBase60.6 g 

SDS10 g 

2.Mix well and adjust the volume to 500 ml withultrapure water.

3.Store at +4°C. The buffer is stable for 6 months when stored at +4°C. 

4.For electrophoresis, dilute this buffer to 1X with water (see page 15). The pH of 

the 1X solution is 8.24. Do not use acid or base to adjust the pH.

NuPAGE® Transfer Buffer

25  mM Bicine

25 mMBis-Tris (free base) 

1 mM EDTA 

pH 7.2 

1.To prepare 125 ml of 20 X NuPAGE® Transfer Buffer, dissolve the following 

reagents to 100 ml ultrapure water: 

Bicine10.2 g 

Bis-Tris (free base)13.1 g 

EDTA0.75 g 

2.Mix well and adjust the volume to 125 ml withultrapure water.

3.Store at +4°C. The buffer is stable for 6 months when stored at +4°C. 

4.For western transfer, dilute this buffer to 1X with water (see page 32). The pH 

of the 1X solution is 7.2. Do not use acid or base to adjust the pH.

NuPAGE® LDS 

Sample Buffer 

106 mMTrisHCl

141 mMTris base 

2% LDS 

10% Glycerol 

0.51 mM EDTA 

0.22 mM SERVA® Blue G250 

0.175 mM Phenol Red 

pH 8.5 

1.To prepare 10 ml of 4 X NuPAGE® LDS Sample Buffer, dissolve the following 

reagents to 8 ml ultrapure water: 

TrisHCl0.666 g 

TrisBase0.682 g 

LDS0.800 g 

EDTA0.006 g 

Glycerol4 g 

SERVA® Blue G250 (1% solution)0.75 ml 

Phenol Red (1% solution)0.25 ml 

2.Mix well and adjust the volume to 10 ml with ultrapure water.

3.Store at +4°C. The buffer is stable for 6 months when stored at +4°C. 

The pH of the 1X solution is 8.5. Do not use acid or base to adjust the pH.

Tris-Glycine

Native Sample 

Buffer 

100 mMTrisHCl

10% Glycerol 

0.0025% Bromophenol Blue 

pH 8.6 

1.To prepare 10 ml of 2 X Tris-Glycine Native Sample Buffer, mix the following 

reagents: 

4 M TrisHCl4 ml 

10% Glycerol2 ml 

0.1% BromophenolBlue0.5 ml 

DeionizedWater3.5 ml 

2.Mix well and adjust the pH of the solution is 8.6.

3.Store at +4°C. The buffer is stable for 6 months when stored at +4°C. 

4.Use this buffer to prepare samples for non-denaturing NuPAGE® Tris-Acetate

gel electrophoresis 

Tris-Glycine

Native Running 

Buffer 

25 mMTris base 

192 mMGlycine

pH 8.3 

1.To prepare 1000 ml of 10 X Tris-Glycine Native Running Buffer, dissolve the 

following reagents in 900 ml deionized water: 

TrisBase29 g 

Glycine144 g 

2.Mix well and adjust the volume to 1000 ml with ultrapure water.

3.Store at room temperature. The buffer is stable for 6 months when stored at 

room temperature. 

4.For native electrophoresis, dilute this buffer to 1X with water (see page 16). The 

pH of the 1X solution is 8.3. Do not use acid or base to adjust the pH.