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Optimize the dilution of the dye provided with PROTEOSTAT® Aggresome detection kit (ENZ-51035) to improve staining. Also incubationb time can be increeased id the dye staining is too low. Confirm that no changes in the protocols for cell permeabilization are made if they have been previously optimized with the same sample type. If a diffuse background signal is detected, there might be some residual detergent. Introducing additional wash steps after permeabilization might help in improving the results. Additional troubleshooting suggestions can be found in pages 18-19 of product manual. Do not hesitate to reach out to Enzo techn support if you need to discuss your results in details.