Thermo Fisher Scientific Slide-A-Lyzer Concentrating Solution User guide

Brand: Thermo Fisher Scientific

Model: Slide-A-Lyzer Concentrating Solution

Type: User guide

INSTRUCTIONS

Pierce Biotechnology

PO Box 117

(815) 968-0747

www.thermoscientific.com/pierce

3747 N. Meridian Road

Rockford, lL 61105 USA

(815) 968-7316 fax

Number Description

66528 Slide-A-Lyzer Concentrating Solution, 200mL with 10 resealable plastic bags (3” × 4”), for use with

0.5mL and 3mL capacity Slide-A-Lyzer Dialysis Cassettes

66529 Slide-A-Lyzer Concentrating Solution, 460mL with 10 resealable plastic bags (5” × 8”), for use with

12mL and 30mL capacity Slide-A-Lyzer Dialysis Cassettes

66530 Slide-A-Lyzer Concentrating Solution, 25mL with one resealable plastic bag (3” × 4”), for use with

a 0.5mL and 3mL capacity Slide-A-Lyzer Dialysis Cassette

Storage: Upon receipt store concentrating solution at room temperature.

Introduction

During dialysis, many samples increase in volume and dilute a valuable protein sample. The Thermo Scientific™

Slide-A-Lyzer™ Concentrating Solution used with a Thermo Scientific Slide-A-Lyzer Cassette can quickly reduce sample

volume, returning the sample to its original concentration or further concentrating it to a desired volume. Concentration is

achieved by diffusion, drawing water and other small molecules out of the cassette and into the solution contained in a plastic

bag. The volume reduction rate is comparable to centrifugation methods for sample concentration.

Using the Slide-A-Lyzer Dialysis Cassette, dialysis and concentration can occur in the same device, alleviating potential

protein loss when switching from one device to another. Additionally, unlike other available products, the Slide-A-Lyzer

Concentrating Solution is composed of a high molecular weight compound that cannot traverse the dialysis membrane and

contaminate the sample.

Important Product Information

• If pH control is critical for the sample, use a sample buffer concentration of at least 100mM. The low buffer salts in the

sample will equilibrate between the sample and Concentrating Solution, reducing their concentration in the sample. For

example, when using a 3mL sample in 100mM phosphate and 25mL of Concentrating Solution, the phosphate will be

diluted to approximately10mM.

• Do not use concentrating solution with 20K Slide-A-Lyzer Cassettes.

Procedure for Sample Concentration

1. Determine the amount of concentrating solution and resealable bag size to use based on the Slide-A-Lyzer Cassette size

being used (Table 1).

Table 1. Concentrating solution volume and bag size to use.

Cassette

(mL)

Concentrating Solution

(mL)

Resealable Bag

Size

0.1-0.5

3”

4”

0.5-3

3”

4”

3-12

5”

8”

12-30

5” × 8”

0631.5

66528 66529 66530

Slide-A-Lyzer Concentrating Solution

Pierce Biotechnology

PO Box 117

(815) 968-0747

www.thermoscientific.com/pierce

3747 N. Meridian Road

Rockford, lL 61105 USA

(815) 968-7316 fax

2. Use a graduated cylinder to measure the required amount of concentrating solution and pour it into the appropriate

resealable bag. Drain the graduated cylinder into the resealable bag for 1 minute

3. Place the Slide-A-Lyzer Cassette containing the sample into the concentrating solution inside the resealable bag. For the

12mL cassette fold the bag as indicated in Figure 1.

to allow the cylinder to empty.

4. Lay the bag containing the cassette on the counter, allowing the solution flow to cover the bottom membrane of the

cassette. Use the palm of your hand to push the concentrating solution over the top membrane of the cassette, and

continue upward to remove the air from the bag. Make sure there are no air bubbles between the cassette membrane

windows and the concentrating solution. Seal the bag.

5. For the 12 and 30mL cassettes, fold the resealable bag as indicated in Figures 1 and 2. Place the bag on a rocking

platform. Set a rocking speed so that the cassette within the bag is moving slightly (~35 RPM). As an alternative, the bag

containing the concentrating solution can be kneaded periodically.

Note: Any mixer that provides enough motion to keep the cassette moving slightly can be used, including nutating and

microplate mixers.

Figure 1. Folded reasealable bag containing

a 12mL Thermo Scientific Slide-A-Lyzer

Cassette. Bag is folded approximately in

thirds.

Figure 2. Folded reasealable bag

containing a 30mL Thermo

Scientific Slide-A-Lyzer Cassette.

6. Carefully monitor the volume decrease as concentration will occur rapidly (see Figures 3-5).

Note: Wear gloves for the next 2 steps.

7. When the sample reaches the desired volume, remove the cassette from the bag.

8. To avoid introducing concentrating solution into the sample, wash the outside of the cassette extensively with deionized

water. Also wash the cassette cavity by performing the following steps:

• Fill a 5-10mL syringe with deionized water. Partially insert needle into the needle port without touching or

penetrating the gasket located within the cassette.

• Flush the cassette cavity to eliminate any residual concentrating solution that could contaminate the sample.

• Continue to flush until the water can be seen in the cavity flowing freely away from the needle port area. The water

flushing action can be observed through the transparent cassette.

9. Remove sample from the cassette using a clean syringe and needle, being careful not to puncture the membrane.

0.5

1.5

2.5

3.5

020 40 60 80 100 120

Time (minute)

Recovered Volume (ml)

10K

3.5K

050 100 150 200

Time (minute)

Recovered Volume (ml)

12 ml

30 ml

Figure 3. Concentration rate for a 3mL Slide-A-Lyzer Cassette.

Figure 4. Concentration rate for 12 and 30mL Slide-A-Lyzer Cassettes.

Pierce Biotechnology

PO Box 117

(815) 968-0747

www.thermoscientific.com/pierce

3747 N. Meridian Road

Rockford, lL 61105 USA

(815) 968-7316 fax

0.1

0.2

0.3

0.4

0.5

0.6

020 40 60

Time (minute)

Recovered Volume (ml)

0.5 ml

Figure 5. Concentration rate for a 0.5mL Thermo Scientific Slide-A-Lyzer Cassette. Because the 0.5mL

cassette has a small membrane area, there can be significant variation in concentration time.

Procedure for Sample Concentration in Slide-A-Lyzer Mini Units

Use a 2.0 or 1.5mL centrifuge tube for each sample to be concentrated. Make sure the Slide-A-Lyzer Mini Unit fits inside the

microcentrifuge tube.

1. Add 1.4mL Concentrating Solution to a 2mL microcentrifuge tube or 1.1mL into a 1.5mL tube.

Note: The volume of Concentrating Solution required may vary slightly with tubes from different manufacturers.

Determine the volume needed with water before adding the Concentrating Solution.

2. Using a pipette, add sample to the bottom of the unit.

3. Wearing gloves, place the unit into the centrifuge tube containing the Concentrating Solution. Avoid trapping air bubbles

under the membrane when inserting the unit into the tube.

4. Allow concentration to occur at room temperature until desired volume is reached.

5. Wearing gloves, use a pipette to retrieve the sample.

100

120

020 40 60 80 100 120

Time (minutes)

Recovered Volume (ml)

1.5 ml tube

2.0 ml tube

Figure 6. Concentration rate for Thermo Scientific Slide-A-Lyzer

MINI Units using 1.1mL of Concentrating Solution in a 1.5mL

microcentrifuge tube and 1.4mL of Concentrating Solution in a 2.0mL

microcentrifuge tube.

Pierce Biotechnology

PO Box 117

(815) 968-0747

www.thermoscientific.com/pierce

3747 N. Meridian Road

Rockford, lL 61105 USA

(815) 968-7316 fax

Slide-A-Lyzer Dialysis Cassette Product Numbers and Descriptions

Membrane Molecular-weight Cutoff (MWCO)

2000

Cassette

Size 3500 7000 10,000

0.1-0.5mL* 66205 (10-pk)

66333 (10-pk)

66335 (Kit) 66373 (10-pk)

66375 (Kit)

66383 (10-pk)

66385 (Kit)

66454 (GI)

0.5-3mL 66203 (10-pk) 66330 (10-pk)

66332 (Kit) 66370 (10-pk)

66372 (Kit)

66380 (10-pk)

66382 (Kit)

66455 (GI)

3-12mL 66212 (8-pk) 66110 (8-pk)

66107 (Kit) 66710 (8-pk)

66707 (Kit)

66810 (8-pk)

66807 (Kit)

66453 (GI)

12-30mL 66230 (6-pk) 66130 (6-pk) NA 66830 (6-pk)

66456 (GI)

Kits include package of cassettes, plus an equal number of float buoys, syringes and needles.

GI = Gamma (γ) Irradiated package of cassettes.

NA = Not Available.

*2K MWCO cassettes in this size are best used for 0.2-0.5mL samples.

Products are warranted to operate or perform substantially in conformance with published Product specifications in effect at the time of sale, as set forth in

the Product documentation, specifications and/or accompanying package inserts (“Documentation”). No claim of suitability for use in applications regulated

by FDA is made. The warranty provided herein is valid only when used by properly trained individuals. Unless otherwise stated in the Documentation, this

warranty is limited to one year from date of shipment when the Product is subjected to normal, proper and intended usage. This warranty does not extend to

anyone other than Buyer. Any model or sample furnished to Buyer is merely illustrative of the general type and quality of goods and does not represent that

any Product will conform to such model or sample.

NO OTHER WARRANTIES, EXPRESS OR IMPLIED, ARE GRANTED, INCLUDING WITHOUT LIMITATION, IMPLIED WARRANTIES OF

MERCHANTABILITY, FITNESS FOR ANY PARTICULAR PURPOSE, OR NON INFRINGEMENT. BUYER’S EXCLUSIVE REMEDY FOR NON-

CONFORMING PRODUCTS DURING THE WARRANTY PERIOD IS LIMITED TO REPAIR, REPLACEMENT OF OR REFUND FOR THE NON-

CONFORMING PRODUCT(S) AT SELLER’S SOLE OPTION. THERE IS NO OBLIGATION TO REPAIR, REPLACE OR REFUND FOR PRODUCTS

AS THE RESULT OF (I) ACCIDENT, DISASTER OR EVENT OF FORCE MAJEURE, (II) MISUSE, FAULT OR NEGLIGENCE OF OR BY BUYER,

(III) USE OF THE PRODUCTS IN A MANNER FOR WHICH THEY WERE NOT DESIGNED, OR (IV) IMPROPER STORAGE AND HANDLING OF

THE PRODUCTS.

Unless otherwise expressly stated on the Product or in the documentation accompanying the Product, the Product is intended for research only and is not to

be used for any other purpose, including without limitation, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses, or

any type of consumption by or application to humans or animals.

Current product instructions are available at www.thermoscientific.com/pierce. For a faxed copy, call 800-874-3723 or contact your local distributor.

Thermo Fisher Scientific Slide-A-Lyzer Concentrating Solution User guide

Thermo Fisher Scientific Slide-A-Lyzer Concentrating Solution User guide

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