Capture the total protein stain-free blot image before adding any chemiluminescent substrate to the blot. This image will be used as the normalization channel.
▶ 00:14
2
Select normalization channel
Open the analysis toolbox and select lane and bands. Use the normalization channel dropdown to add the total protein stain-free blot image as the normalization channel alongside the chemi image.
▶ 00:26
3
Configure band detection settings
When the band detection window opens, set the detection sensitivity based on signal strength. For strong signals, select low detection sensitivity and confirm to allow Image Lab to automatically detect lanes and bands.
▶ 01:01
4
Select normalization reference lane
Choose one of the total protein lanes as the normalization lane. This lane will serve as the reference for calculating normalization factors across all samples.
▶ 01:39
5
Adjust and verify background subtraction
Select the total protein blot and click adjust background on the toolbar. Review the gray section underneath the red line across all lanes to ensure consistent background subtraction, adjusting the subtraction scroll bar if needed.
▶ 01:56
6
Review normalization results
Close the lane profiler and click the analysis table button to view detailed results including band volume, intensity before normalization, normalization factors, and adjusted volumes for each lane.
▶ 02:40
7
Export normalized data to Excel
Click the export to Excel analysis button to save the complete normalization results and normalized protein data for further analysis or reporting.