b'Pierce Reversible Protein Stain Kits for Nitrocellulose or PVDF MembranesA great alternative to Ponceau SThermo Scientific Pierce Reversible Protein Stain Kits for either nitrocellulose or PVDF membranes provide a rapid and sensitive alternative to Ponceau S stain for protein detection on nitrocellulose or PVDF membranes after transfer from polyacrylamide gels (Figures 11 and 12). The lower limit of detection with this method is 2550 ng per band (at least 5x more sensitive than traditional Ponceau S staining). The simple and quick staining protocol results in turquoise-blue bands that do not fade and are easily photographed for future reference. The stain can be easily reversed in less than 15 minutes. Subsequent western blot detection is unaffected because the stainA1 2 3 4 B1 2 3 4does not alter the protein and is completely removed. The treated membrane does not interfere with conventional chemiluminescent or chromogenic detection using horseradish peroxidase (HRP) and alkaline phosphatase (AP) substrates. In addition, the stain is compatible with N-terminal sequence analysis of proteins excised and eluted from the membrane.Features: Pierce Reversible Protein Stain Ponceau S stain Better than Ponceau Smore sensitive, easier toFigure 11. Pierce Reversible Protein Stain and Ponceau S stain: document, permanent until reversed (Table 11) a comparison of staining of bacterial lysate with overexpressed GST protein, on nitrocellulose. Increasing amounts of lysate wereSensitivehigh-avidity, total-protein stain; lower limit ofseparated on two 420% Tris-glycine polyacrylamide gels and transferred to nitrocellulose membranes. (A) One blot was treated with Pierce detection equals 25 to 50 ng per band Reversible Protein Stain for 30 seconds and destained according to the protocol. (B) Another blot was stained with 0.1% Ponceau S stainSpecificdetects only protein; does not bind or interactfor 5 minutes and destained. The blot stained with Pierce Reversible with other electrophoresis or sample components Protein Stain demonstrates superior visual detection of bands. GST lysate loading volumes: lane 1: 5 L, lane 2: 10 L, lane 3: 15 L. Lane 4: 10 LRapidstains in less than 5 minutes (nitrocelluloseMW marker.membranes) or in less than 30 minutes (PVDF membranes); erase and reverse staining in less than 15 minutes Stablecomponents are stored at room temperature to help save refrigerator space and eliminate equilibration stepsLearn more at thermofisher.com/proteinstains26'