Thermo Fisher Scientific Pierce Western Blot Transfer Buffer 10X Methanol-free User guide

Brand: Thermo Fisher Scientific

Model: Pierce Western Blot Transfer Buffer 10X Methanol-free

Type: User guide

INSTRUCTIONS

Pierce Biotechnology

PO Box 117

(815) 968-0747

www.thermo.com/pierce

3747 N. Meridian Road

Rockford, lL 61105 USA

(815) 968-7316 fax

Number Description

35040 Pierce Western Blot Transfer Buffer, 10X, Methanol-free, 5L

35045 Pierce Western Blot Transfer Buffer, 10X, Methanol-free, 500mL

Storage: Upon receipt store at 4°C. Product is shipped at room temperature.

Introduction

The Thermo Scientific™ Pierce™ Western Blot Transfer Buffer is for transferring proteins from SDS-PAGE gels to

nitrocellulose or PVDF membranes. This transfer buffer is compatible with tank and semi-dry transfer units and is

specifically formulated to be used without methanol and without chilling. This buffer is supplied as a 10X concentrate and is

diluted with deionized water before use.

Procedure for Tank or Semi-Dry Transfer of Proteins

A. Additional Materials Required

• Wet (tank) or semi-dry protein transfer apparatus and power supply

• Protein transfer sponges (for tank transferred only)

• Filter paper

• Transfer membrane

B. Transfer Proteins from Gel to Membrane

1. Dilute the Pierce Western Blot Transfer Buffer to 1X with deionized water.

2. Equilibrate transfer sponges, filter paper and membrane in 1X transfer buffer for 10-15 minutes with gentle rocking.

Note: PVDF membranes must be pre-wet with methanol before equilibrating in transfer buffer. For semi-dry transfer,

use two pieces of thin filter paper (~1.25mm) or one piece of ultra-thick filter paper (~2.5mm) on top and bottom.

3. After electrophoresis, equilibrate gel in ultrapure water for 5-10 minutes with gentle agitation.

4. Equilibrate gel in 1X transfer buffer for 5-10 minutes with gentle agitation.

5. Assemble tank transfer sandwich between cathode and anode in a tank transfer unit (with ice pack) as indicated below.

Assemble semi-dry transfer sandwich directly on anode plate of semi-dry transfer unit as indicated below.

Note: Eliminate air bubbles between gel and membrane with roller or clean pipette.

6. Connect the tank transfer unit to the power supply and apply a continuous current of 400mA for 1 hour.

Connect the semi-dry transfer unit to the power supply and apply a continuous voltage of 25V for 15-60 minutes.

7. Remove and rinse membrane with ultrapure water. Proceed with protein detection method.

Tank Transfer Sandwich

Semi-Dry Transfer Sandwich

1. Cathode (-)

2. Sponge

3. Filter Paper

4. Gel

5. Membrane

6. Filter Paper

7. Sponge

8. Anode (+)

1. Cathode (-)

2. Filter Paper (2 sheets)

3. Gel

4. Membrane

5. Filter Paper (2 sheets)

6. Anode (+)

2171.3

35040 35045

Pierce Western Blot Transfer Buffer,

10X, Methanol-free

Pierce Biotechnology

PO Box 117

(815) 968-0747

www.thermo.com/pierce

3747 N. Meridian Road

Rockford, lL 61105 USA

(815) 968-7316 fax

Troubleshooting

Problem

Possible Cause

Solution

Inefficient transfer Salt build-up on electrodes Thoroughly wash tank electrodes or electrode surfaces

with deionized water after each use

Membrane, filter paper, or gel

insufficiently equilibrated in transfer

buffer

Equilibrate membrane filter paper, and gel in 1X

transfer Buffer for 10-15 minutes before transfer

Insufficient transfer time Increase transfer time from 1 hour to 75 minutes

PVDF membrane was not pre-wet with

methanol Pre-wet PVDF membrane with methanol and then

equilibrate for 10-15 minutes in 1X transfer buffer

Inconsistent transfer Air bubbles trapped between gel and

membrane When assembling sandwich use a roller or pipette to

remove air bubbles between the gel and the membrane

Related Thermo Scientific Products

88600 Western Blotting Filter Paper, 8cm × 10.5cm, 100 sheet/box

77012 Nitrocellulose Membrane, 0.2μm, 8cm × 12vm, 25/pkg.

88013 Nitrocellulose Membrane, 0.2μm, 7.9cm × 10.5cm, 15/pkg.

88024 Nitrocellulose Membrane, 0.2μm, 8cm × 8cm, 1 roll

88018 Nitrocellulose Membrane, 0.45μm, 30cm × 3.5m, 1 roll

77010 Nitrocellulose Membrane, 0.45μm, 8cm × 12cm, 25/pkg.

88014 Nitrocellulose Membrane, 0.45μm, 7.9cm × 10.5cm, 15/pkg.

88025 Nitrocellulose Membrane, 0.45μm, 8cm × 8cm, 15/pkg.

22860 Low-Fluorescence PVDF Transfer Membrane, 0.2μm, 7cm × 8.4cm, 10/pkg.

88518 PVDF Transfer Membrane, 0.45μm, 26.5cm × 3.75m, 1 roll

88585 PVDF Transfer Membrane, 0.45μm, 10cm × 10cm, 10 sheets

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.

subsidiaries. Printed in the USA.

Thermo Fisher Scientific Pierce Western Blot Transfer Buffer 10X Methanol-free User guide

Thermo Fisher Scientific Pierce Western Blot Transfer Buffer 10X Methanol-free User guide

Thermo Fisher Scientific Pierce Western Blot Transfer Buffer 10X Methanol-free User guide

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