![]() If you are using an Odyssey DLx or an Odyssey M, it is best to place the silicone mat on top of the membrane. Image Membraneĭo not allow the membrane to dry during imaging. If you wish to visualize several HKPs in the 700 nm and 800 nm channels, you can do so by destaining Revert according to the instructions in your pack insert. You do not need to destain Revert 700 Total Protein Stain in this protocol as you will be visualizing your HKP and OLI in the 800 nm channel. Using approximately 5 mL of Revert 700 Wash Solution (P/N 926-11012), rinse the membrane two times for 30 seconds at room temperature with gentle shaking.ĭecant wash solution thoroughly, then briefly rinse the membrane with ultrapure water. Incubate the membrane in 5 mL of Revert 700 Total Protein Stain solution for 5 minutes at room temperature with gentle shaking.įor more information about the Revert protocol, see /revert.ĭecant total protein stain solution thoroughly. Stain membrane with Revert 700 Total Protein Stain. Without clearance, staining may be uneven. Revert 700 Total Protein Stain will cause the membrane to swell. Then rinse in TBS or PBS (no detergent) for 5 minutes at room temperature with gentle shaking.ĭo not allow the membrane to dry from this point on.īefore moving to the next step, ensure the membrane container provides a minimum clearance of 1/8th of an inch on all sides. Rehydrate the membrane after fully drying.įor nitrocellulose membranes, incubate the membrane in TBS or PBS (no detergent) for 5 minutes at room temperature with gentle shaking.įor PVDF membranes, first rehydrate using 100% methanol for 30 seconds. Overnight at room temperature as a stopping point. Place the membrane on top of a piece of clean filter paper and allow it to dry: Add methanol to the stain reagents as indicated on each bottle.Īfter transfer is complete, fully dry the membrane.Stain with Revert™ 700 Total Protein Stain Use the steps below for detection of a target in the 800 nm channel. Method 1: Single-Color Western Blot (800 nm target only) The total protein normalization method using Revert 700 Total Protein Stain provides a quick and reliable method for normalizing target Suitable for Western blot normalization adds significant time and cost Validating housekeeping proteins that are In Western blot analysis, normalization is essential for accurate, reproducibleĬomparison of protein levels. This will allow you to determine if there are any irregularities that indicate you should run the blot again to get more robust results. With a total protein stain, you will be able to monitor protein transfer across the entire blot at all molecular weights. You can also remove the total protein stain and process the blot to detect two different targets in the 700 and 800 nm channels. With an imaging system capable of multiplex detection, you can image the total protein stained with Revert 700 Total Protein Stain in the 700 nm channel and image a target protein in the 800 nm channel on the same blot. Stained proteins produce signal that can be detected visibly and by imagers capable of detecting dyes that have excitation and emission maxima near 700 nm. Revert 700 Total Protein Stain provides high signal intensity with a broad, linear quantitative range. ![]() Revert 700 Total Protein Stain is a membrane stain that fluoresces at 700 nm and does not covalently modify sample proteins and therefore does not affect antibody binding or quantification. Revert Destaining Solution: 0.1 M sodium hydroxide, 30% (v/v) methanol, in Wash Solution: 6.7% (v/v) glacial acetic acid, 30% (v/v) methanol, in.Shelf Life: 6 months from date of receipt.Sufficient reagent for up to 4 mini blots. Revert 700 Total Protein Stain and Wash Solution Kit Sufficient reagent for up to 50 mini blots. Revert 700 Total Protein Stain Kit, 250 mL Sufficient reagent for up to 20 mini blots. Revert 700 Total Protein Stain Kit, 100 mL
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