4X Protein Loading Buffer

Specifications

  • Size: 15 mL

  • Storage: Store at ambient temperature in a tightly sealed container.

    Do not store Protein Loading Buffer with β-mercaptoethanol for more than two weeks.

  • Shelf Life: One year from date received.

Formulation

125 mM Tris-HCl, pH 6.8
50% Glycerol
4% SDS
0.2% (w/v) Orange G

Applications

4X Protein Loading Buffer is a loading buffer for protein gel electrophoresis. This loading buffer is not visible when imaged on an Odyssey Fc, Odyssey XF, Odyssey CLx, or Odyssey DLx and is only faintly visible when imaged in the visible channels on the Odyssey M. The buffer is optimized for use with SDS-PAGE and Tris-glycine-SDS running buffer.

Protocol

Before use, warm buffer slightly until any SDS crystals disappear. SDS crystals may form at temperatures lower than 28 °C.

Protocol for 4X Protein Loading Buffer

  1. Add 100 μL of β-mercaptoethanol to 900 μL of 4X Protein Loading Buffer and mix well.

  2. For every 3 μL of sample, add 1 μL of protein loading buffer. The recommended concentration for electrophoresis is 1X.

  3. Follow your usual protocol for denaturing protein samples.

Protocol for 2X Protein Loading Buffer

  1. Combine 475 μL of 4X Protein Loading Buffer with 475 μL of ultrapure water, then

    add 50 μL of β-mercaptoethanol. Mix well.

  2. For every 1 μL of sample, add 1 μL of protein loading buffer.

  3. Follow your usual protocol for denaturing protein samples.