1300 Electrophoresis 2006, 27, 1300–1302

Satoshi Sugawa Short Communication

Tatsuzo Kawano
Yukinori Nara
Tetsu Isomura High-throughput sodium dodecyl sulfate
ZoeGene Corporation polyacrylamide gel electrophoresis by linking
Yokohama, Japan
conventional gel plates with automated liquid
Received July 27, 2005 handler via Gel Adaptor
Revised September 24, 2005
Accepted September 29, 2005 The automation of SDS-PAGE required substantial investment. To lower this barrier, we
devised a cost-effective, simple, and general method where samples are loaded on
SDS-PAGE gels held by a newly devised “Gel Adaptor” on a general automated liquid
handler. In this method, the liquid handler loads samples on the SDS-PAGE gels held at
an angle of 807 on the Gel Adaptor so that the samples can be vertically loaded on the
narrow paths of the wells of the slanted gels. This method allows the automated liquid
handler to load 144 samples on SDS-PAGE gels in approximately 10 min, greatly re-
ducing the time and cost for high-throughput SDS-PAGE analyses.

Keywords: Automated liquid handler / Automatic loading / Automation / High-

throughput / SDS-PAGE DOI 10.1002/elps.200500548

The automation of SDS-PAGE required substantial
investment for a high-spec instrument, e.g., a custom-
made instrument for SDS-PAGE, or an instrument for CE
that separates proteins similarly to SDS-PAGE. Capillary
SDS-DALT electrophoresis is a CE system giving results
similar to SDS-PAGE [1]. The gel in the capillary however
consists of less branched polymers than those for typical
SDS-PAGE because of the physico-chemical restraints in
the capillary. The sample proteins have to be fluorescently
labeled to enhance the detection sensitivity through the
short lightpath of the capillary. For these reasons, the
separation pattern of the proteins of the CE may not be
consistent with that of conventional SDS-PAGE. There is
also a need to perform Western blotting following SDS-
PAGE in many cases, while it is not possible with capillary
SDS-DALT electrophoresis.
To make high-throughput SDS-PAGE more approachable
to the broader population of protein researchers, it would
be convenient if a large number of samples were auto-
matically loaded by general laboratory robotics because
sample loading is one of the most laborious and time-
consuming steps when performing high-throughput SDS-
PAGE. E-Gel® (Invitrogen) is an agarose gel plate with a
base of 96-well plate format that is placed on a stage for
Correspondence: Satoshi Sugawa, ZoeGene Corp., 6-50-8, Sekido,
Tamashi, Tokyo 206-0011, Japan
E-mail: ssugawa@nyc.odn.ne.jp
Fax: 181-42-337-1124
96-well plates on an automated liquid handler and is
loaded with DNA samples automatically [2]. Since the
probes of the liquid handler are directing vertically in the
normal settings, it was difficult to apply this method to
loading samples on SDS-PAGE gels because the thick-
ness of either the probe or the disposable tip is usually
greater than the gap between the plates holding the
polyacrylamide gel. We solved this problem by holding
the gel plate at a slant angle using the “Gel Adaptor” so
that the vertical probe could load samples on the wells of
the gel without the steric hindrance. Here we report this
cost-effective, simple, and general method in which SDS-
PAGE gels are held on an automated liquid handler by the
Gel Adaptor, allowing 144 samples to be loaded in ap-
proximately 10 min.
As shown in Fig. 1a, the Gel adaptor was designed to hold
eight slab gel plates at an angle of 807. In order for tips to
dispense samples precisely, this slant position was
devised (Fig. 1b, right) because the width of the tips is
greater than the gap between the front and the back gel
plates (Fig. 1b, left) and the distance from the top to the
gel was too long (22 mm) to directly aim to the sample
wells uprightly (Fig. 1b, center). The slanting angle was
determined by two restriction factors: (i) it should be
smaller than the taper angle of the tips (Fig. 1c, left); (ii) it
should be sufficiently great so that the gel plate at the
edge of the adaptor does not obstruct the space of the
adjacent stage (Fig. 1c, right). The condition for (i) was
,857 and that for (ii) was .707 with the liquid handler that

© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.electrophoresis-journal.com

Electrophoresis 2006, 27, 1300–1302
Figure 1. (a) Plan of the Gel Adaptor. (b) Three ways of
sample loading. (c) Maximum (left) and minimum (right)
possible gel angles.
we employed. We therefore adopted 807 as the feasible
gel plate angle for the Gel Adaptor. The longitudinal and
lateral distances of the bottom of the Gel Adaptor were
designed to be the same as those of a standard 96-well
plate (85 mm and 128 mm, respectively) so that it can fit a
typical stages for 96-well plates installed in general liquid
handlers.
The Gel Adaptor was manufactured according to the plan
as shown in Fig. 1a. Biomek FX (Beckman-Coulter,
Fullerton, CA, USA) was used as the automated liquid
handler in this study. Span-8 (a block of eight independ-
ent probes) of Biomek FX was used for loading samples
on plural gels simultaneously to reduce the time for the
procedure. Pure™ BioRobotix™ tips (catalog no. 919-261,
Molecular BioProducts, San Diego, CA, USA) were
used as disposable tips. e-PAGEL® (model E-R10L,
© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
General 1301
10% acrylamide gel, 18 wells; ATTO Corp., Tokyo, Japan)
was used as a pre-cast gel and Page-Run (model AE-
6531P, ATTO Corp.) was used as the apparatus for SDS-
PAGE. Molecular weight marker “DAIICHI” III (Daiichi
Pure Chemicals, Tokyo, Japan) was used as samples for
SDS-PAGE.
The operational procedures are as follows: (i) set SDS-
PAGE gel plates on the Gel Adaptor; (ii) set the Gel
Adaptor on the automated labware positioner (ALP),
which is a stage for 96-well plates on Biomek FX; (iii) fill
sample wells of the gels with SDS-PAGE running buffer;
(iv) start automatic loading of the samples; and (v) manu-
ally transfer the gel plates to the electrophoresis appara-
tus and start running SDS-PAGE.
To operate Biomek FX for the automatic loading, Biomek
FX-controlling-software was used as described in the
manual. X-, Y-, and Z-axis’ positions of the upper left
sample well seen from the top were registered as 2.7 mm,
2.5 mm, and 89 mm, respectively. The intervals of
18 wells along the X-axis were registered as 4.5 mm and
those of eight wells along the Y-axis were registered as
9 mm.
As shown in Fig. 2, all the samples were accurately and
reproducibly dispensed in the wells. When the automatic
loading was performed without filling the wells with SDS-
PAGE running buffer prior to the loading, the air bubbles in
the wells hampered the samples from gravitationally set-
tling down to the bottoms of the wells (data not shown),
showing that the prior buffer filling is an essential step for
this method. The time required for loading 144 samples
(18 samples per gel, eight gels) was approximately
10 min. When eight 12-well gels were put on the Gel
Figure 2. Example of the result of SDS-PAGE performed
by the automatic loading method.
www.electrophoresis-journal.com
1302 S. Sugawa et al.
Adaptor so that the positions of the wells matched those
of 96 wells in a 96-well plate, 96-channel liquid handler
was applied to this automatic loading, reducing the load-
ing time by ,1/10 (data not shown). Considering the time
and labor of conventional manual sample loading, this
automatic loading is advantageous for high-throughput
SDS-PAGE analyses.
The other advantage for this automatic loading is that a
complicated format of sample loading can be conducted
without error once it is programmed. The record of the
loading format can also be traced back whenever
needed.
We designed the Gel Adaptor to fit in the stages for the
standard 96-well plates, so this technique can be
employed by most of the automated liquid handlers on
the market. Researchers, who already have those instru-
ments, can introduce the high-throughput SDS-PAGE
system into their laboratories with the Gel Adaptors,
which are much easier and cheaper to obtain than a cus-
tomized automated SDS-PAGE system.
© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Electrophoresis 2006, 27, 1300–1302
The Gel Adaptor links conventional gel plates for SDS-
PAGE slantedly with an automated liquid handler, making
it capable of loading samples into the sample wells in the
gels automatically. With this method, 144 samples can be
loaded on eight gels in approximately 10 min. Because
the Gel Adaptors are designed to fit in any automated
liquid handlers having stages for 96-well plates, this
method should be available to a broad population of
researchers, and is expected to greatly reduce the time
and cost for high-throughput SDS-PAGE analyses.
This work was supported by ZoeGene Corporation. We
thank Dr. T. Shibui for the critical reading of the manu-
script.
References
[1] Hu, S., Jiang, J., Cook, L. M., Richards, D. P. et al., Electro-
phoresis 2002, 23, 3136–3142.
[2] E-Gel® Technical Guide, Version C, 25-0645, Invitrogen, 18
February 2005.
www.electrophoresis-journal.com