Western Blot: What happens to my protein?

Monitoring and Normalization of Sample Protein

The immunochemical detection of specific proteins by Western Blot analysis is performed by different steps including: protein extraction, separation, transfer, blocking, antibody incubation, washing, target detection.

The protein transfer rate is less than 100%. Transferred proteins are not covalently bound to the blotting membrane. Therefore, the amount protein of each sample is continuously decreased (see figure below). The loss of protein is caused by different sources (e.g. hydrophobicity, mechanical shearing forces,...).

Valid Normalisation of Western Blots

The patented Smart Protein Layers (SPL) technology offers you the easy but sensitive detection of sample protein  during all stages of the Western Blot procedure. The precise detection of sample amount present on the blot at the same time the target is immunodetected allows for the accurate normalisation of target protein expression.

Patented SPL Technology

The corresponding SPL kit comes with fluorescent SPL label which rapidly bind to the sample protein prior electrophoresis. Depending on sample properties two different workflows can be used.

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SPL EASY workflow
Very easy usage of the SPL kit  limited to samples within a range of 25μg - 75μg of extracted total protein.

SPL ADVANCED workfow
Advanced usage of the SPL kit for scarce samples and/ or particular buffer compositions. This approach provides further information about the sample, the labeling reaction and monitors the complex WB workflow.

Both the easy and advanced workflow offer precise protein transfer monitoring and allow for experiment-to-experiment comparisons.