Plan, run, diagnose and document Western blots
Make the next Western blot decision with evidence, not guesswork.
Choose the task in front of you. Each resource separates a reasonable starting condition from the observation that should make you change it.
Choose the job you need to finish
Complete protocol
Run the full workflow with decision points for sample preparation, gels, transfer, antibodies and detection.
2Plan an experiment
Define expected size, controls, extraction, membrane, detection range and the smallest useful pilot.
3Troubleshoot interactively
Answer four observations and receive a prioritized checking order, missing evidence and one next action.
4Review diagnostic cases
Work through evidence, competing causes and the next discriminating experiment for common failure patterns.
5Calculate volumes
Calculate loading, sample mixes, direct and intermediate antibody dilutions, TBST and gel-range considerations.
6Record the experiment
Use editable, locally saved and printable experiment and troubleshooting worksheets.
7Prepare buffers
Use clearly labeled starting formulations and compatibility boundaries.
8Check the evidence
Open manufacturer protocols, methodological papers and reporting guidance.
Start from what you see on the membrane
Open the full troubleshooting page →The whole blot is blank
Check acquisition and detection, then transfer, then the positive control and antibody chain.
Target is absent but control is present
Check biology, expected size, sample fraction and target-specific transfer before titrating antibody.
Uniform dark background
Check exposure and secondary dilution before changing several blocking and wash conditions.
Extra bands or a smear
Separate sample overload or degradation from true isoforms, processing and cross-reactivity.
Uneven or patchy signal
Use the position of the artifact to inspect bubbles, contact, coverage, contamination and imaging surface.
Use the handbook across the whole experiment
01
Before loading
Use planning and the record sheet to confirm target, sample, controls, expected size and detection range.
02
During the run
Use the protocol to verify observable evidence for lysis, separation and transfer.
03
At imaging
Save the whole membrane, multiple exposures and unsaturated raw data—not only the best-looking image.
04
After failure
Use the assistant and troubleshooting record; change one variable that distinguishes competing hypotheses.
Tools that turn guidance into a repeatable workflow
Diagnostic case library
Break an appearance into observation, evidence, competing causes and a discriminating experiment.
Printable records
Keep actual conditions, raw-file locations and the next single variable on one page.
Pipetting-aware calculators
Go beyond arithmetic by checking negative volumes, pipetting limits and intermediate dilutions.
Use the site as a decision aid, not a universal SOP
Conditions depend on the target, antibody, sample, gel, membrane and imager. Begin with product-specific validation, preserve source data, and change one causal block at a time.