Welcome to the Empiria Studio® Software Help!
Explore the infographic below to learn about Empiria Studio.
A Project is used to store related Experiments. An Experiment can be either a Validation or a Target Analysis.
Tip: Keep related Validations and Target Analyses together in a Project to ensure your data are organized when it’s time to publish.
Validations are required to ensure your Quantitative Western blot results will accurately reflect the changes you are trying to measure.
Validate that your primary antibody is both specific and selective to the target antigen.
Determine a combined linear range for your target and internal loading control. Accurate quantification requires that your internal loading control and target both be detected within the same linear range.
Total protein stain and target
Determine the range of sample loading that produces a linear signal response for both the target and the total protein stain (the combined linear range).
Housekeeping protein and target
Determine the range of sample loading that produces a linear signal response for both the target and a housekeeping protein (the combined linear range).
Housekeeping protein validation
Show evidence that your chosen housekeeping protein is stably expressed.
Quantify housekeeping protein signal and compare to reference protein signal. If the housekeeping protein signal varies but the reference protein signal is constant, the housekeeping protein should not be used for normalization.
Housekeeping protein validation with a total protein stain
Quantify and compare housekeeping protein signal to total protein stain signal. If the housekeeping protein signal varies compared to the total protein stain, the housekeeping protein should not be used for normalization.
Once you have completed the appropriate Validations, begin the Analysis workflow that corresponds to the internal loading control you used for normalization.
Normalization using a total protein stain
Quantify your total protein stain and target. Empiria Studio will normalize the target signal to the total protein stain signal.
Normalization using a housekeeping protein
Analyze your target bands, using your HKP for normalization.
Normalization using a post-translation modification
Analyze your modified target protein, using the pan-protein for normalization.
Compare replicate Target Analyses to get statistically significant results and your final answer.
Create a Publication pack to share multiple Experiments with publishers and colleagues. Validations are valuable supplementary data and can be included as well.