Zymography Analysis Software | Phoretix 1D
Automate the quantification of gelatin and casein zymograms. Phoretix 1D detects lysis bands, corrects for the inverted zymogram profile, and gives you reproducible protease and MMP activity data in minutes, not hours.
Trusted by the world’s leading pharmaceutical, industrial and academic laboratories.
Powerful technique. Hard to quantify consistently
A zymogram is the inverse of a normal gel. Active proteases digest the gelatin or casein substrate, so your signal appears as clear zones on a darkly stained background. Measuring those cleared bands by eye, or wrestling with a general-purpose tool that expects dark bands on a light background, is slow and introduces the operator-to-operator variability that undermines a method already described as only semi-quantitative. Phoretix 1D is purpose-built to remove that variability.
Built for the zymogram, not adapted to it
Handles the inverted profile
Clear lysis bands on a stained background are the opposite of a standard gel. Invert the lane profile in one click so cleared zones are measured correctly as signal, never mistaken for background.
Background subtraction for uneven staining
Rolling-ball and other background methods correct patchy Coomassie staining and gel-to-gel variation, per lane and per channel, so activity values stay comparable across runs.
Automated lane and band detection
Next-generation automatic detection finds lanes and lysis bands in seconds, even on distorted or wonky gels, with full manual control wherever you want it.
What you can quantify
- Gelatin zymography: MMP-2 and MMP-9 (gelatinase A and B), the most common application, central to cancer invasion, inflammation and neurovascular research.
- Casein zymography: serine proteases and calpains, applied from oncology through to calpain and calpastatin activity in food and meat science.
- Reverse and in-gel zymography: quantify protease-inhibitor activity and other substrate-embedded assays with the same workflow.
From image to activity data in four steps
- Import your zymogram. Open TIFF, PNG, JPEG, BMP or IMG straight from any gel doc system. 16-bit TIFF recommended.
- Detect lanes and bands. Automatic lane and lysis-band detection in seconds, with manual add, split and remove when you want it.
- Invert and subtract background. One click handles the clear-band profile; choose a background method to correct uneven staining.
- Quantify and report. Reproducible band volumes exported to CSV, clipboard or a formal PDF report for your ELN or LIMS.
Why labs choose Phoretix 1D for zymography
- Reproducible results across operators, labs and imaging devices
- Device agnostic: Bio-Rad, Cytiva, Azure, Thermo Fisher, Syngene, Vilber
- Full analysis in under a minute per image
- Handles high-definition, multiplexed and 16-bit images
- Automatic and manual control on a per-lane, per-channel basis
- 21 CFR Part 11 ready with AuditSafe for regulated QC
Used across the protease-activity literature
- Oncology and metastasis: MMP activity drives tumor invasion and migration, a core readout in cancer-progression studies.
- Neuroscience and vascular biology: MMP-9 release in stroke and blood-brain-barrier research, where reproducible quantification matters clinically.
- Food and dairy science: casein zymography for calpain and calpastatin activity, linked to meat tenderness and post-mortem processing.
- Microbiology and environment: profiling microbial and environmental proteases across gelatin and casein substrates.
- Pharmaceutical QC: protease and gelatinase activity assays run under 21 CFR Part 11 with full traceability.
- Inflammation and immunology: gelatinase and serine-protease activity in inflammatory and immune-response models.
Zymography under 21 CFR Part 11
Combined with AuditSafe, Phoretix 1D delivers secure sign-ins, granular permissions, audit logs, electronic signatures and image authenticity checks, from captured zymogram to final report. Full in-person IQ/OQ validation packages are available for regulated environments.
Zymography analysis FAQs
Can Phoretix 1D handle the clear bands of a zymogram?
Yes. Zymogram bands are cleared zones on a stained background, the inverse of a normal gel. Phoretix 1D inverts the lane profile in one click so those cleared zones are measured correctly as signal rather than background.
Does it support gelatin and casein zymography?
Yes, plus reverse zymography. Any substrate-embedded assay that produces banded proteolysis on an imaged gel can be analyzed with the same workflow.
Which imagers and image formats are supported?
Any of them. Phoretix 1D is device agnostic and reads TIFF, PNG, JPEG, BMP and IMG files from any gel documentation system, with 16-bit TIFF recommended for best results.
Is software-based zymography quantification accurate?
Zymography is semi-quantitative by nature. Software does not change that, but it removes the operator-to-operator variability of manual measurement and applies consistent background correction, which substantially improves reproducibility.
Can I use it in a regulated GMP environment?
Yes. Combined with AuditSafe, Phoretix 1D meets 21 CFR Part 11, EU Annex 11 and GMP requirements, and full IQ/OQ validation packages are available.
See it on your own zymograms
Download a free trial and run Phoretix 1D on your own gelatin or casein zymograms. We will send your trial licence within one business day.