Large Volume Tris Buffer Saline and Tween 20 (20X)
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Description: Tris Buffered Saline and Tween 20 are in a 20X concentration for use as a wash buffer for immunolabeling techniques. Tween 20 is added to promote more effective washings, resulting in decreased non-specific background staining. Tween 20 also aids reagent spreading on slides.
Diluting Procedures: To make 1X TBS/Tween 20 solution dilute conctentrate 20X with distilled water. Use 50ml PBS concentrateper 950ml DI water. Measure and pour appropriate volume of 20X PBS concentrate into a mixing flask and add DI water to final volume. Stir briefly. The 1X solution should be pH 7.6+-0.2 The pH of the soution may require slight adjustment. Adjust pH with 1N HCL(if pH is high) or 1N NaOH (if pH is low).
Storage: Store Tris Buffered Saline/Tween 20 at room temperature. Concentrate is stable for 24 months from date of manufacture. Diluted solution should be used within one week. if solution becomes cloudey, discard and make fresh solution. Note: The salts in the 25X solution are highly concentrated. Temperatures below 44 degrees F (6 degrees C) may cause salts to precipitate. If this occus, stir the solution at RT for about 30 mins to redissolve the salts (use an alcohol cleaned stir bar to prevent bacterial contamination.
Step-by-Step Instructions: Note pH of buffer at 20X is approximately 7.2. After diluting to 1X, pH should be 7.4. Depending on pH of water, the pH of the solution may require pH adjustment. Pour 20X TBS in a mixing flask and add DI water to final volume. Stir briefly. 60 ml of concentrate yields 1.2 liters, and 125 ml yields 2.5 liters.
