Western Blotting is Easy with Pierce Protein ... - Fisher Scientific

Western Blotting is Easy with Pierce Protein 

research products from Fisher Scientific 

StEP 1: SDS-PAGE 

Separate protein sample by electrophoresis. 

• Pierce SDS-PAGE Sample Prep Kit 

• Precise Protein Gels 

• Tris-Hepes-SDS Running Buffer 

• Lane Marker Reducing Sample Buffer 

• Lane Marker Non-Reducing Sample Buffer 

• Pierce Protein Molecular Weight Markers 

StEP 2: Electro-transfer 

Transfer proteins to membrane. 

• Pierce Reversible Protein Stain Kit for 

Nitrocellulose Membranes and 

for PVDF Membranes 

• Tris-Glycine Transfer Buffer 

• Pierce Western Blot Signal Enhancer 

• Pierce Antibody Extender NC 

• Nitrocellulose Membranes, 0.2 µm and 0.45 µm 

• PVDF Membrane, 0.45 µm 

• Low-fluorescence PVDF Membrane, 0.2 µm 

• Western Blotting Filter Paper 

StEP 3: Blocking 

Block nonspecific sites. 

• Protein-free Blocking Buffer 

• StartingBlock Blocking Buffers in PBS or 

TBS with or without 0.05% Tween ® -20 

• SuperBlock ® Blocking Buffers in PBS or TBS 

with or without 0.05% Tween-20 

• Casein in PBS and in TBS 

• BSA in PBS and in TBS 

StEP 4: Formulate Wash Buffers 

Choose a buffer. 

• Phosphate Buffered Saline 

• Tris Buffered Saline 

• Tween-20 

†See patent information on reverse page. 

Part of Thermo Fisher Scientific 

For detection of proteins that cannot be efficiently transferred to a 

membrane, thermo Scientific Pierce In-Gel Detection technology 

allows positive identification of proteins directly in a gel. 

For convenience and economy, we also offer complete Western blotting kits 

that include chemiluminescent substrates, enzyme-conjugated antibodies, 

blocking buffers and standard buffers. 


StEP 5: Primary and Secondary Detection Reagents 

Incubate the membrane with antibody. 

For a complete list, visit the antibody 

selection guide on our website. 

For direct detection methods we offer: 

• Monoclonal Antibodies 

• Fluorescent Probes and Labeling Kits 

• Enzyme Labeling Kits 

For indirect detection methods we offer: 

• Biotinylation Kits 

• Protein A, Protein G and Protein L labeled with fluorescein, 

rhodamine, HRP, AP or biotin 

• Avidin, Streptavidin and NeutrAvidin Biotin-Binding Protein labeled 

with fluorescein, rhodamine, HRP or AP 

• Clean-Blot IP Detection Reagents 

• Secondary antibodies labeled with fluorescein, rhodamine, 

HRP, AP or biotin 

• DyLight Secondary Antibody and Streptavidin Conjugates 

StEP 6: Enzyme Substrates 

Add the detection reagent. 

Chemiluminescent Substrates: 

• Pierce ECL Substrate 

• SuperSignal West Pico Chemiluminescent Substrate 

also available in an economical 1-L package 

• SuperSignal West Femto Maximum Sensitivity Substrate 

• SuperSignal West Dura Extended Duration Substrate 

• Lumi-Phos WB Substrate for Alkaline Phosphatase 

Colorimetric Substrates: 

• Pierce Chloronaphthol 

• TMB-Blotting 

• NBT/BCIP 

• Metal Enhanced DAB 

StEP 7: Film 

Expose the membrane to X-ray film. 

• CL-XPosure Film 5 x 7” sheets; 8 x 10” sheets; 9.5 x 11.8” sheets; 

18 x 24 cm sheets and 35 x 43 cm sheets 

• Pierce Background Eliminator Kit 

StEP 8: Stripping Buffer 

Reprobe the blot if necessary. 

• Restore Western Blot Stripping Buffer 

• Restore PLUS Western Blot Stripping Buffer for 

High-Affinity Antibodies 

See overleaf for part numbers and prices 

For Customer Service, call 031 - 352 32 00 

To fax an order, use 031 - 352 32 50 

To order online: www.fishersci.se 

Email: fisher.se@thermofisher.com 

HRP 

SuperSignal ® 

Substrate 

terms and Conditions 

Please note that all sales are subject 

to our standard terms and conditions. 

Full details can be found on the inside 

back cover of our catalogues or visit 

our website www.fisher.co.uk

Free Western Blotting Handbook and 

troubleshooting Guide 

This recently updated 73-page handbook 

contains full-length and abbreviated 

Western blotting protocols, tips on 

antibody and blocking buffer optimisation, 

a chemiluminescent substrate selection 

guide, a procedure for stripping and 

reprobing Western blots, tips on clearer 

film development, and other valuable 

information. 

Request your free copy today! 

Download it from www.fishersci.se/ 

Product # Description Pkg. Size Price 

224-019060 Nitrocellulose Membrane, 0.2 µm 15/pkg 1.179:- 

7.9 cm x 10.5 cm 

224-019080 Nitrocellulose Membrane, 0.45 µm 1 roll 3.915:- 

30 cm x 3.5 m 


7.9 cm x 10.5 cm 


8 cm x 8 cm 


8 cm x 12 cm 


8 cm x 8 cm 


8 cm x 12 cm 

224-005590 Low-Fluorescence PVDF 10/pkg 1.008:transfer 

Membrane, 0.2 µm 7 cm x 8.4 cm 

224-019280 PVDF transfer Membrane, 0.45 µm 10 sheets 1.125:- 

10 cm x 10 cm 

224-019260 PVDF transfer Membrane, 0.45 µm 1 roll 3.051:- 

26.5 cm x 3.75 m 

224-019290 Western Blotting Filter Paper 100 sheets 486:- 

Blocking Buffers 


224-012420 StartingBlock (PBS) Blocking Buffer 1 L 2.115:- 

A protein-based blocker formulation in phosphate-buffered 

saline (pH 7.5) for use in Western blotting and ELISA applications. 

224-012440 StartingBlock (tBS) Blocking Buffer 1 L 2.115:- 

A protein-based blocker formulation in Tris-buffered saline 

(pH 7.5) for use in Western blotting and ELISA applications. 

224-012430 StartingBlock t20 (PBS) Blocking Buffer 1 L 2.114:- 

A protein-based blocker formulation in phosphate-buffered 

saline at pH 7.5 with 0.05% Tween-20 Detergent and Kathon ® 

Antimicrobial Agent. 

224-012450 StartingBlock t20 (tBS) Blocking Buffer 1 L 2.114:- 

A protein-based blocker formulation in Tris-buffered saline 

at pH 7.5 with 0.05% Tween-20 Detergent and Kathon 

Antimicrobial Agent. 

Contact Information 

224-012300 SuperBlock (PBS) Blocking Buffer Belgium and Europe, 1 L 1.980:- 

the Middle East 

and Africa Distributors 

Tel: +32 53 85 71 84 

224-012310 France 

Tel: 0 800 50 82 15 

SuperBlock t20 (PBS) Blocking Buffer 1 L 2.034:- 

(Contains 0.05% Tween-20 Detergent) 

The Netherlands 

Tel: 076 50 31 880 

Germany 

Tel: 0228 9125650 

United Kingdom 

224-012390 SuperBlock (tBS) Blocking Buffer 

Tel: 0800 252 185 

Switzerland 

Tel: 0800 56 31 40 

Email: perbio.euromarketing@thermofisher.com 1 L 2.178:www.thermo.com/perbio 

United States 

Tel: 815-968-0747 or 800-874-3723 

224-012400 SuperBlock t20 (tBS) Blocking Buffer Customer Assistance E-mail: 

Pierce.CS@thermofisher.com 

www.thermo.com/pierce 

(Contains 0.05% Tween-20 Detergent) 

1 L 2.034:- 

Secondary Antibodies 

Product # Description Pk Size Price 

224-009410 Goat Anti-Mouse IgG (H+L) Peroxidase Conjugated 2mL 2.781:- 

224-009610 Goat Anti-Rabbit IgG (H+L) Peroxidase Conjugated 2mL 2.349:- 

DyLight Conjugates: Package size for these items is 1 mg at 1 mg/ml. 

DyLight DyLight DyLight DyLight DyLight 

Description 488 Dye 549 Dye 649 Dye 680 Dye 800 Dye 

Goat Anti-Mouse IgG (H+L) 224-012070 224-012080 224-012100 224-012110 224-012120 

Goat Anti-Rabbit IgG (H+L) 224-012130 224-012140 224-012160 224-012170 224-012180 

Streptavidin 224-004490 224-004500 224-004520 224-004530 224-004540 

NeutrAvidin Protein 224-005470 224-005480 224-005500 224-005510 224-005540 

Substrates 

224-011300 Pierce ECL Western Blotting Substrate kit 500mL 2.079:- 

224-011330 Pierce ECL Western Blotting Substrate kit 250mL 1.629:- 

224-011310 Pierce ECL Western Blotting Substrate kit 50mL 648:- 

224-011900 SuperSignal West Pico Chemiluminescent Substrate 500mL 2.790:- 

224-011880 SuperSignal West Pico Chemiluminescent Substrate 1L 4.518:- 

224-011850 SuperSignal West Dura Extended Duration Substrate 100mL 4.131:- 

224-012010 SuperSignal West Femto Maximum Sensitivity Substrate 100mL 4.599:- 

224-012050 Lumi-Phos WB Chemiluminescent Substrate 100mL 2.934:- 

Film and Stripping Buffers 

224-011960 CL-XPosure Film, 13 x 18 cm 100/pkg 1.368:- 

224-011980 CL-XPosure Film, 13 x 18 cm 25/pkg 576:- 






224-003330 Pierce Background Eliminator Kit 1.611:- 

Sufficient reagents to prepare 3L working solution 

224-003300 Restore Western Blot Stripping Buffer 500mL 1.611:- 

Sufficient for stripping 25 (8 cm x 10 cm) blots 

224-003320 Restore Western Blot Stripping Buffer 30mL 486:- 

Sufficient for stripping one (8 cm x 10 cm) blot 

224-013590 Restore PLUS Western Blot Stripping Buffer 30mL 486:- 

Sufficient reagent to strip one to two (8 cm x 10 cm) blots 

224-013600 Restore PLUS Western Blot Stripping Buffer 500mL 1.638:- 

Sufficient reagent to strip 25 (8 cm x 10 cm) blotsv 

Thermo Scientific SuperSignal Technology is protected by U.S. patent # 6,432,662. Thermo Scientific Pierce Direct Detection of Biomolecules Technology is protected by U.S. patent # 7,112,411. 

U.S. patents pending on Thermo Scientific Pierce Western Blot Signal Enhancer Technology. Kathon ® is a trademark of Rohm & Haas Company. LumiPhos is a trademark of – and is sourced from – Lumigen, Inc. 

Alexa Fluor ® is a trademark of Invitrogen Corporation. Tween ® is a trademark of ICI Americas Inc. CyDye is a trademark of GE Healthcare. 


1601725 12/08 Printed in the U.S. 

© 2008 Thermo Fisher Scientific Inc. All rights reserved. 

These products are supplied for laboratory or manufacturing 

applications only. Unless indicated otherwise on the inside 

back cover, all trademarks are property of Thermo Fisher 

Scientific Inc. and its subsidiaries. 

Pierce ® Protein Research Products 

2009 Catalog 

Thermo Scientific Pierce 

Protein Research Products 

2009 Catalog 

Cell Lysis & Protein Extraction 

Affinity Purification 

Dialysis and Desalting 

Protein Assays 

Electrophoresis and Staining 

Western Blotting, ELISA 

and Cell Imaging 

Protein Interactions 

Mass Spec, HPLC & GC Reagents 

Protein Labeling 

Crosslinking and 

Protein Modification 

Antibody Production 

and Purfication 

Target-Specific Assays 

and Reagents 

To discuss your exact requirements please contact Fisher Scientific 

For Customer Service, call 031 - 352 32 00 

To fax an order, use 031 - 352 32 50 

To order online: www.fishersci.se 

Email: fisher.se@thermofisher.com 

Download your copy today 

www.fishersci.se/ 

terms and Conditions 

Please note that all sales are subject 

to our standard terms and conditions. 

Full details can be found on the inside 

back cover of our catalogues or visit 

our website www.fisher.co.uk

Western Blotting is Easy with Pierce Protein ... - Fisher Scientific

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