Suspending the Substrate Powders in Buffer

PROCEDURE FOR SUSPENDING THE SUBSTRATE POWDERS IN BUFFER

Materials Required

1.     Substrate Powder, e.g., Elastin-Rhodamine, 200-400 Mesh.

2.     Buffers: 0.2 M Tris pH 8.8 containing 0.01% Trixton* X-100 | 0.2 M Tris pH 8.8

3.      Whatman No. 41 paper or equivalent
 

4.     Powder Funnel

5.     Magnetic Stirrer

 

Procedure

1.    Add the substrate (20mg/ml) to gently stirring buffer containing Triton X-100 and stir until all particles are wetted. Wash the substrate on Whatman No. 41 filter paper with buffer (not containing Triton X-100) until the filtrate is colorless. Resuspend the substrate in buffer (not containing Triton X-100) to 20mg/ml.

NOTE: always maintain a layer of buffer over the substrate while washing. Do no allow the substrate to filter to dryness or clumping of the substrate particles may occur. 
 

Storage: Store at 5┬║C
*Triton is a registered trademark of Rohm and Hass Company.