CORNING Spin-X UF 500 Dane techniczne i instrukcja obsługi - Strona 2

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Introduction

Spin-X

UF concentrators are disposable, single use only ultrafiltration devices

with polyethersulfone membranes (PES) for the concentration and/or purification

of biological samples. Spin-X UF 500 concentrators are suitable for sample volumes

of 100 to 500 µL. Thevertical membrane design and thin channel filtration chamber

minimizes membrane fouling and provides high speed concentrations, even with

particle laden solutions.

Storage Conditions and Shelf Life

Spin-X UF concentrators should be stored at room

temperature. The devices should be used before the

expiration date printed on the box.

Chemical Compatibility

Spin-X UF concentrators are designed for use with

biological fluids and aqueous solutions. For chemical

compatibility details, refer to Table 3 (page 5).

Centrifugal Operation

Spin-X UF 500 concentrators can be used in a benchtop

fixed angle rotor, accepting 2.2 mL centrifuge tubes. In a

single spin, solutions can be concentrated approximately

100-fold.

Required Equipment

1. Centrifuge with fixed angle rotor (minimum 40°) that

can fit 2.2 mL (11 mm diameter) conical bottom tubes.

2. Fixed or variable volume pipettors with gel loading tips

are recommended for sample addition and removal.

Operation

1. Select the most appropriate membrane cut-off for your sample. For maximum

recovery select a molecular weight cut off (MWCO) at least 50% smaller than the

molecular size of the species of interest.

2. Fill concentrator with up to a maximum volume of 500 µL as shown in Table 1

(Ensure lid is fully seated).

3. Insert assembled concentrator into a centrifuge with a fixed angle rotor. Minimum

rotor angle is 40º. Angle concentrator so that the printed window faces upwards/

outwards.

Corning Spin-X UF 500 –

100 to 500 µL capacity

4. Centrifuge at speeds up to 15,000 xg.

5. Once the desired concentration is achieved, remove assembly and recover sample

from the bottom of the concentrate pocket with a pipettor and gel loading tip.

See Table 2 (page 4) for a guide to concentration times. The filtrate tube can be

sealed for storage.

Desalting/Buffer Exchange

1. Concentrate sample to desired level.

2. Empty filtrate container.

3. Refill concentrator with an appropriate solvent.

4. Concentrate the sample again and repeat the process until the concentration of

the contaminating microsolute is sufficiently reduced. Typically, 3 wash cycles will

remove 99% of initial salt content.

Helpful Hints

1. Flow Rate

Filtration rate is affected by several parameters, including MWCO, porosity, sample

concentration, viscosity, centrifugal force and temperature. Expect significantly longer

spin times for starting solutions with over 5% solids. When operating at 4°C, flow

rates are approximately 1.5 times slower than at 25°C. Viscous solutions such as 50%

glycerin will take up to 5 times longer to concentrate than samples in a predominantly

buffer solution.

2. Prerinsing

Membranes fitted to Spin-X UF concentrators contain trace amounts of glycerin and

sodium azide. Should these interfere with analysis they can be removed by rinsing fill

volume of buffer solution or deionized water through the concentrator. Decant filtrate

and concentrate before processing sample solution. If you do not want to use the pre-

rinsed device immediately, store it in the refrigerator with buffer or water covering the

membrane surface. Please do not allow the membrane to dry out.

3. Sterilization of Polyethersulfone Membranes

Polyethersulfone membranes should not be autoclaved as high temperatures will

substantially increase membrane MWCO. To sterilize, use a 70% ethanol solution

or sterilizing gas mixture.