PREPARATION OF REAGENT DYE SOLUTION - (1.320 G/L)
(USING 1 LITER REAGENT DYE CONCENTRATE P/N 30-0601)

Carefully pour the contents of the Reagent Dye Concentrate bottle into the 10 Liter Calibrated Carboy container. Rinse the dye bottle and cap bottle and cap many times with warm distilled water into the Carboy container.

Carefully add 600 ml of Glacial Acetic Acid (use proper safety precautions) to the Carboy container. Cap container and swirl contents to help dissolve dye crystals.

Carefully add distilled water to the Carboy container and bring the level up to the 8 Liter mark. Cap the container and mix contents several times. Let the contents sit for several hours (optimum time is >24 hours) before testing for proper concentration. To help dissolve the crystals in the container, occasionally stir or shake the contents during the waiting period. The reason for the waiting period is to make sure all of the dye crystals are dissolved thoroughly.

After the waiting period is completed, verify the concentration of the Reagent Dye in the Carboy. Verification procedure (Working Reference):

A. Dispense 120 ml or weigh out 121.11 g (" .24g) of the Reagent Dye from the Carboy into the 240 ml Volumetric Flask (PN 35-0527). Add distilled water to the flask and bring the level to the line (bottom of the meniscus); cap and mix several times. This solution will be used to verify for proper concentration in the Carboy.

B. Set the UDY Colorimeter switch to the CAL. position, add distilled water to the Cuvet and adjust to read out .000 on the display. Add standard Reference Dye (PN 30-0613) to the Cuvet (clear water by flushing 2 times with Standard Reference Solution) and adjust using the AGAIN@ knob until a read-out of 0.660 is achieved. The UDY Colorimeter will now give concentration values for the Reagent Solution in Step A. NOTE: ALL SOLUTIONS MUST BE AT THE SAME ROOM TEMPERATURE THROUGH THE UNIT (IDEAL TEMPERATURE IS 25C).

C. After Step B is finished, add the Diluted Reagent Dye Test Solution (25-30 drops) from Step A into the Cuvet and record the reading. Since the Carboy level at this time is only 8 liters, the concentration should be >1.320 g/L or a readout higher than .660. Rule of Thumb: If the reading is higher than .660, add 1.5 ml of distilled water per Liter of Solution for each .001 that the reading is too high. If the reading is lower than .660, add 0.2 ml of Reagent Dye Concentrate (PN 30-0602 - 100 ml bottle) per Liter of Solution for each .001 low. Mix solution after adding dye or water; then retest the Solution. Adjust the solution as needed to get into the ideal range.

THE IDEAL RANGE OF THE CARBOY SOLUTION FOR

PROTEIN TESTING SHOULD HAVE A CONCENTRATION

FROM 0.656 TO 0.664 G/L

When the concentration of the Carboy Solution is in the proper range, transfer the contents into the Dispensing Carboy. Recheck occasionally; and adjust as needed.

Storage and Use of Dye Solutions

Made From Concentrates and Stored In Glass Containers

Reagent and Reference Dye Concentrates may be stored for several years, because water lost through the Container walls is replaced when the dye is diluted to its proper volume for use. Once Reagent or Reference Dye has been made to its final concentration, care must be exercised to prevent concentration changes through water loss by evaporation. To maintain proper concentrations, the following points must be followed:

Store solutions not in use, in tightly capped GLASS containers. Water evaporates from solutions stored in plastic containers (including the Reagent Dye Dispensor).

Before opening containers or using dropper bottles after several hours of non-use, invert the bottle once or twice. This remixes the water droplets which form above the liquid level. The Reagent Dye Solution in the Dispensor should be stirred to remix the droplets formed in it.After opening storage or dropper bottles, wipe off both the bottle neck and inside of cap with a tissue before pouring solution.

Rewipe bottle necks with a tissue after solutions are poured before recapping. This prevents dye accumulation and crystallization. Serious errors can result if the dye crystallizes in the bottle threads and later mixes in the solutions. Keep bottle capped when not in use B including vent on Reagent Dye Dispensor.

Solutions not used from a dropper bottle within a week or two should be discarded. If a full bottle will not be used within this time, only partially fill the bottle. Always discard remaining solution before adding fresh solution.

Prevent formation of air bubbles in the cuvet by squeezing dropper bottles gently and directing the flow against the side of the cuvet.

Since evaporation losses from the Reagent Dye Dispensor cannot be prevented, Reagent Dye concentration must be checked at least monthly.

When properly stored and handled, prepared dye solutions (in glass containers) will remain at the correct concentration for over one year. Because the Standard Reference Dye Solution is the standard against which all other concentrations are measured, these instructions are especially important for it.