Lonza Rockland, Inc.
www.lonza.com
biotechserv@lonza.com
Tech Service: 800-521-0390
Customer Service: 800-638-8174
Document # 18652-1007-03
Rockland, ME 04841 USA

GelBond® Film
Casting Agarose Gels on GelBond Film
I. Introduction
GelBond® Film is a transparent, flexible plastic film
designed to support agarose gels. This convenient,
ready to use film has a specially treated hydrophilic sidethat firmly adheres agarose gels as they form. Gels
remain adhered to the film from the time they are castthrough all electrophoresis or immuno-diffusion, fixing,
staining, and destaining procedures.

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GelBond® Film provides many advantages over the useof glass plates alone. Gels cast on it are easy to handle
and can be dried and stored permanently. GelBond®
Film often eliminates the need for photography. The useof GelBond® Film is particularly suitable for gels stainedwith Coomassie® Brilliant Blue or agarose silver stain.
Agarose gels can be silver stained only after drying onto
GelBond® Film.

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GelBond® Film is non-porous and can not be used withelectroblotting or capillary blotting applications.

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GelBond® Film is intended for use with agar and agarose
gels. GelBond® Film is recommended for most agarose
gel applications including gel diffusion,electroimmunodiffusion, serum protein electrophoresis,
isoelectric focusing and nucleic acid electrophoresis.

Cover the GelBond® Film with a sheet of blotting
paper or the interleaving paper supplied with theGelBond® Film and firmly roll with a rubber roller orwipe with tissues to squeeze out any air bubbles
and excess fluid from behind the GelBond® Film.
Carefully wipe off any excess liquid at the edges.
The GelBond® Film should now be tightly adhered tothe glass plate.
Arrange the side spacers and the bottom spacers on
top of the GelBond® Film.
Place the top plate over the bottom plate.
Tape the sides of the cassette with separate piecesof tape.
Tape the bottom of the cassette with a separate
piece of tape. This allows the tape on the bottom tobe removed for electrophoresis without disturbing
the tape at the sides of the gel.
Clamp the plates together.

Casting a vertical agarose gel
1. Prepare agarose solution according to themanufacturer’s instructions and cool to 60ºC.
2. Pre-warm the assembled cassette and a 60 ml
syringe for 15 minutes by placing in a 55ºC oven or
by using a heat gun.
3. Pour agarose into a pre-warmed 60 ml syringe fittedwith a 16-gauge needle.
4. Wedge the needle tip between the plates in theupper corner of the cassette with the needle opening
directed toward the back plate.
5. Angle cassette and inject the agarose solution at amoderate, steady rate allowing the agarose to flow
down one side spacer.
6. Fill until the agarose solution goes just above theglass plates.
7. Insert one end of the comb, then slowly insert therest of the comb until the teeth are at an even depth.
10. Insert the comb into the agarose to the minimal
depth necessary to accommodate your samples.
11. Place extra clamps on the side of the glass plates tohold the comb in place.
12. Cool the gel at room temperature for 15 minutes.
13. Place the gel at 4ºC for 20 minutes.

GelBond® PAG Film is available for use withpolyacrylamide gels.

II. Vertical Cassette Casting
The casting cassette consists of two glass plates,
GelBond® Film, spacers, and clamps.
Assemble the cassette as follows:
1. Spread a few drops of distilled water or 0.1%
nonionic detergent solution on the back plate.
2. Lay a sheet of GelBond®
3. Film, cut slightly smaller (1 mm-2 mm) than theglass plate, onto the plate with the hydrophilic sideup (water droplets spreadon the hydrophilic side but bead up on thehydrophobic side).

114. Remove the clamps at the top of the gel.
15. Remove excess agarose with a scalpel or razor
blade.
16. Squirt running buffer in the spaces between thecomb and the gel.
17. Slowly and gently lift the comb straight up. Allow airor buffer to enter the well area to release thevacuum which forms between the agarose and thecomb.
18. Flush wells with running buffer.
19. Store gels overnight in a humidity chamber or in asealed bag with a buffer-dampened paper towel.

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Lower the gel on the wetted area. Avoid trapping air
under the GelBond® Film. Wipe excess fluid fromthe edges of the film.
Blot the surface of the gel briefly with a sheet of finegrained blotting paper.
Apply samples by using either a sample application
mask or filter paper sample applicator pieces.
Run the electrophoretic separation.

V. Fixing and Staining

General instructions for fixing and staining
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Preparing for electrophoresis
NOTE: Since agarose does not adhere well to glass,leave as many clips in place as possible. For someelectrophoresis chambers, it is helpful to seal thespacers at the top of the gel with molten agarose.
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Remove the tape at the bottom of the cassette.
Place the cassette into the chamber at an angle tominimize the number of bubbles which can collect inthe well area.
Rinse out well area with a running buffer.

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Disassemble the casting cassette as follows
1. Remove the tape and all clamps.
2. With the cassette lying flat, insert a flat spatula
between the glass plates and twist gently to breakthe seal. Carefully remove the top plate, leaving thegel and the GelBond® Film attached to the backplate.
3. Remove the spacers and lift the gel, attached to theGelBond® Film from the back plate.

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III. Open Casting for Horizontal Isoelectric Focusing
As an alternative to using a casting cassette, gels can beformed by an open casting method. This technique is
optimal for gels of 1.0 mm-1.5 mm thickness. However,there is a greater likelihood of nonuniformity andevaporative loss with this method. Gels must be cast on
a level surface to assure a uniform thickness. Caremust be taken to prevent the agarose solution fromoverflowing the edge of the GelBond® Film and runningunder the support film.
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For open casting, adhere GelBond Film to a glass
plate as described previously.
As with cassette casting prewarm the plate withGelBond® Film attached prior to gel casting.
After casting and once the gel has set, refrigerate
the gel in a humidity chamber for one hour prior touse.

IV. Preparation for Horizontal Electrophoresis
1. Trim the gel with a razor blade to ensure straightedges.
2. Spread a small volume of 0.1% nonionic detergent
or distilled water on the cooling platen or bed of theelectrophoresis chamber.
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Soak the gel in the appropriate fixative for 2
minutes, then gently agitate for 10-20 minutes.
NOTE: Prolonged exposure to TCA fixative candisadhere the gel from the GelBond® Film.
Wet a sheet of fine grained blotting paper with
distilled water and place it on the gel surface.
Overlay the blotting paper with 4-6 layers ofabsorbent paper toweling. Place a glass plate on
top and weight it down with a 1 kg-2 kg weight for
20 minutes.
Remove the weight, glass plate and the paper
toweling. Rewet the blotting paper thoroughly with
distilled water and gently lift it off the gel surface.
Rinse the gel in a large volume of distilled water for
5-10 minutes with agitation to remove residual
fixative (or ampholytes from IEF gels).
To accelerate deproteinization ofimmunoelectrophoresis gels to assure a clear
background, saline soakes and press blotting should
be repeated 2-5 times.
Clamp the GelBond® Film, gel side out, onto a glass
plate to prevent curling during drying. Dry the gelcompletely in a forced hot air oven (55ºC-60ºC) oron the lab bench overnight. Oven drying usuallytakes <30 minutes.
Transfer the dried gel to a stain bath for 10-30
minutes. Float the gel on the surface of the solution
with the gel facing down into the stain. This
prevents precipitated stain from settling on the gel.
Remove the stained gel, rinse it briefly in distilledwater, destain for a few minutes as needed, thenrinse briefly in distilled water.
Clamp the gel to a glass plate, gel side out and dry
the gel in a forced-air oven for approximately
15 minutes or dry at room temperature overnight.
Gels will crack if overdried. The gel can be read,stored or otherwise used without further treatment.

VI. Special Techniques•A. Press Blot Transfer to Membranes
GelBond® Film is neither porous nor electricallyconductive so it cannot be used with conventionalor electrophoretic blotting procedures. However, unfixed
proteins and nucleic acids can be transferred tonitrocellulose or other membranes by direct pressure
blotting. This procedure is nearly identical to the press
blot drying previously described, except that a sample
binding membrane (e.g. nitrocellulose) is substituted forthe sheet of blotting paper and no weight is placed on
the glass plate. Proteins and nucleic acids are drawn
out of the gel and onto the membrane, withapproximately 20% of the proteins transferred after just1½ minutes and up to 85% transferred after 35-40
minute blotting time.

•To uncurl GelBond® Film cut from a roll, place thecut film, rewound in the opposite direction, into abeaker of boiling water for 5 minutes.
Soak curled gels in 0.5%-2% glycerol and re-dry atroom temperature.

Ordering Information
GelBond® Film, 0.2mm thick sheets 100 per package
Catalog No.
Size (mm)53734
85 x 10053745
110 x 125
53746
100 x 150
53748
110 x 205
53749
160 x 18053759
125 x 245
53761
124 x 258

B. Ultraviolet Light and Fluorescent Staining• GelBond® Film exhibits background fluorescencewhen exposed to UV light. This background may
interfere with visual detection of fluorescent bands
such as cardiac isoenzymes or nucleic acids stainedwith ethidium bromide. If photography is used,background fluorescence can be screened out by
using red, orange (e.g. Wratten® Gelatin Filter), andUV camera filters.
• GelBond® Film also blocks much of the light below
300 nm, so nucleic acid gels should be inverted (gelside down) onto the light box in order to expose theseparated bands to full light intensity. Thefluorescent bands photograph well through
GelBond® Film using this method.
• To stain gels backed with GelBond® Film withGelStar®, SYBR® Green or SYPRO® Red Gel Stains,follow the directions for staining vertical gels in thestains instruction sheet.

GelBond® Film, 0.2mm thick rolls, 16.5 meters long
Catalog No.
Roll Width53740
102 mm
53750
127 mm
53760
152 mm
53780
203 mmRelated Products
GelBond® PAG FilmIsoGel® Agarose
Lonza Agarose

VII. TroubleshootingAdhesion• For maximum adhesion cast agarose solutions at60ºC-65ºC.
• Always preheat casting cassette or glass plate withGelBond® Film attached to 60ºC-65ºC.
• Only cast gels on the hydrophilic side of GelBond®
Film.
• Store GelBond® Film in a dry location.
• Prolonged exposure to TCA can disadhere gels fromthe film.
• Include 0.5%-2% glycerol in the final rinse or soak
step to prevent gels from cracking during drying.
• Destain times may have to double to increase theirefficiency.

For Laboratory Use.

GelBond® Film curl
• Avoid high temperature (>65ºC) drying andoverdrying of the gel.
• Always clamp the gel to a glass plate while drying.

©2007 Lonza Rockland, Inc.

Manufactured for Lonza Rockland, Inc.
GelBond and GelStar are trademarks of FMC Corporation.
Wratten is a trademark of Eastman Kodak. SYBR and SYPRO
are trademarks of Molecular Probes, Inc. All other trademarks
herein are marks of the Lonza Group or its affiliates.

All Rights Reserved.
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