Thermo Fisher Scientific Pierce Background Eliminator User guide

Brand: Thermo Fisher Scientific

Model: Pierce Background Eliminator

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

21065 Pierce Background Eliminator

Kit Contents:

Reagent A, 100mL

Reagent B, 100mL

Storage: Upon receipt store at room temperature. Product shipped at ambient temperature.

Introduction

Thermo Scientific™ Pierce™ Background Eliminator allows background to be removed from overexposed film after it has

been developed. This product is the ideal solution for researchers who want to visualize images that may be difficult to

discern because of high background, overexposed bands or speckling without spending valuable time repeating experiments

or trying to time the exposure precisely. Pierce Background Eliminator Solution will produce clear images from

Western/Southern/Northern blots, gel shifts or any other assay that requires film to capture an image (Figure 1). For

applications requiring densitometry measurements, the process reduces signal intensity evenly across the entire film so that

relative densitometry values are consistent.

Pierce Background Eliminator works on freshly developed films or films that have been stored for years. The working

solution can be used at room temperature and does not need to be used in the darkroom. The entire process takes only a few

minutes and may be stopped when the exposure looks just right.

Procedure for Using Pierce Background Eliminator

1. Thoroughly wash developing chemicals from film.

2. Place two containers of deionized water nearby for rinsing film. Containers must be large enough to fully submerge film.

3. Prepare Working Solution immediately before use by adding Reagent A and Reagent B to deionized water in the

amounts indicated below. The Working Solution is stable for up to 30 minutes.

Film Size

Container Size

Water

Reagent A

Reagent B

5” × 7”

8” × 8” × 2”

250mL

10mL

8” × 10”

8”× 10” × 2”

500mL

20mL

4. Add freshly prepared Working Solution to a glass or plastic container large enough to fully submerge film.

5. Place one film in the Working Solution and rock container. Make sure film does not stick to container bottom. Flip film

over every 15-30 seconds.

6. Remove film when desired image appears.

Note: Monitor film closely or desired image may be erased. Erasing background over a light box may help visualize the

progress.

7. Immediately submerge film in the first water rinse for a few seconds. The solution continues to erase until the film is

removed.

8. Remove film from first water rinse and place in second water rinse. Make sure to thoroughly wash film.

9. Allow film to dry.

1339.2

21065

Pierce™ Background Eliminator

Pierce Biotechnology

PO Box 117

(815) 968-0747

www.thermoscientific.com/pierce

3747 N. Meridian Road

Rockford, lL 61105 USA

(815) 968-7316 fax

Troubleshooting

Problem

Cause

Solution

Background is not removed

evenly across the entire film

Film stuck to the container bottom or not

flipped consistently

Rock container during incubation and flip

film every 15-30 seconds

More than one film was incubated in the

Working Solution

Incubate only one film at a time

Image disappears too quickly

Working Solution was too concentrated

Working Solution may be diluted to slow

reaction

Not sure when to stop the

reaction

Difficult to determine when results are

optimal

Stop reaction and photograph data at several

time-points

Note: Film can be repeatedly incubated in

the Working Solution

Working Solution no longer

removes background

Saturation or depletion of Working

Solution

Use more newly prepared Working Solution

Reagent B has darkened with

time

Normal reagent property

Reagent will function properly

Yellow tint on film

Inadequate washing

Thoroughly wash film to remove excess

Working Solution

Extremely dark film saturated or

depleted the Working Solution

Use more newly prepared Working Solution

Use only one piece of film at a time

Related Thermo Scientific Products

34090 CL-XPosure™ Film, 5” × 7” sheets, 100 sheets/pkg

32106 Pierce ECL Western Blotting Substrate, 500mL

34080 SuperSignal West Pico Chemiluminescent Substrate, 500mL

34075 SuperSignal West Dura Substrate, 100mL

34095 SuperSignal West Femto Maximum Sensitivity Substrate, 100mL

33500 Pierce In-Gel Chemiluminescent Detection Kit – Rabbit

33505 Pierce In-Gel Chemiluminescent Detection Kit – Mouse

20148 LightShift™ Chemiluminescent EMSA Kit

Figure 1. Recombinant Human TNFα was separated by SDS-PAGE using a 4-20% gel

and transferred to a nitrocellulose membrane. The membrane was blocked and detected

with mouse anti-Human TNFα followed by Goat anti-Mouse-HRP (Product No. 31434) and

Thermo Scientific™ SuperSignal™ West Dura Substrate (Product No. 34075). The blot was

exposed to film for 30 seconds, resulting in considerable background speckling (A). The film

was then treated with Pierce Background Eliminator for 2 minutes (B).

Pierce Biotechnology

PO Box 117

(815) 968-0747

www.thermoscientific.com/pierce

3747 N. Meridian Road

Rockford, lL 61105 USA

(815) 968-7316 fax

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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

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