Azure Chemiluminescent Western Demo Kit

Workflow

Step 1

Sample preparation

Step 2

Perform electrophoresis

Protein separation by SDS PAGE: Run the electrophoresis using your standard lab conditions, until the dye front reaches the bottom of the gel.

Step 3

Buffer preparation

Step 4

Equilibriate membrane and transfer

Step 5

Dispense 10mL of Buffer C into incubation tray. Incubate membrane for 10 minutes at RT with gentle agitation.

Step 6

Add primary antibody and Buffer D. Incubate for 30 minutes minutes at RT with gentle agitation.

Step 7

Wash membrane 1X for 15 minutes with 25mL of Buffer E with gentle agitation.

Step 8

Prepare Radiance Substrate

Combine Radiance components 1:1 in sufficient amounts to obtain 0.1 mL/cm² of your membrane.

Step 9

Apply Radiance Substrate

Incubate for 5 minutes.

Step 10

Image blot