Home›Biochemistry›Import of ÄKTA chromatograms for purity analysis
Steps
1Export chromatogram from ÄKTA system00:05
2Prepare SDS-PAGE gel with fractions00:35
3Image gel on trans-illuminator01:00
4Analyze gel image with software01:20
5Import chromatogram into analysis software01:37
6Generate combined purity report01:57
BiochemistryCytiva
Import of ÄKTA chromatograms for purity analysis
Protocol
Difficulty
intermediate
Steps
1
Export chromatogram from ÄKTA system
View the purified protein chromatogram on the ÄKTA system and export it as an RC file for downstream analysis.
▶ 00:05
2
Prepare SDS-PAGE gel with fractions
After completing the ÄKTA run, prepare an SDS-PAGE gel using the collected fractions. Stain the gel using the commercial staining standard (Novagen) for band visualization.
▶ 00:35
3
Image gel on trans-illuminator
Visualize the stained SDS-PAGE gel using a calibrated trans-illuminator with a light source that measures optical density (OD) units, the industrial standard for quantifying visible protein stains.
▶ 01:00
4
Analyze gel image with software
Import the gel image into quantitation software and define lanes, background, and bands. Compare profiles across different lanes and runs by overlapping the data.
▶ 01:20
5
Import chromatogram into analysis software
Import the exported ÄKTA chromatogram file into the quantitation software to link the chromatogram peaks with the SDS-PAGE gel analysis results.
▶ 01:37
6
Generate combined purity report
Export a combined report that integrates both the chromatogram from the ÄKTA run and the SDS-PAGE gel analysis data for comprehensive purity documentation.