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High-Throughput Pre-Cast Gel System

  • Protein Separation and Analysis for Increased Sample Throughput

    Description:
    The E-PAGE™ High-Throughput (HTP) Pre-Cast Gel System is the only system available for fast, bufferless, high-throughput protein analysis. It consists of E-PAGE™ pre-cast gels, E-Base™ integrated devices, the E-Holder™ platform, and the free E-Editor™ 2.0 software.

E-PAGE™ Gels

  • E-PAGE™ Gels are self-contained, pre-cast gels that include a gel matrix and electrodes packaged inside a disposable, UV-transparent cassette. Two gel types (Table 1 and 2) are currently available to meet your separation needs:

    • E-PAGE™ 48 8% Gels— each gel contains 48 sample lanes and 4 marker lanes with a 3.2-cm separation length; the E-PAGE™ 48 Gels can be loaded with a 12 or 8-channel pipettor (Figure 1A) or 4- or 8-tip robotic systems*
    • E-PAGE™ 96 6% Gels— each gel contains 96 sample lanes and 8 marker lanes in a patented staggered-well format with a 1.6-cm run length (Figure 2); well openings are compatible with the standard 96-well plate format and can be conveniently loaded with a multichannel pipettor or 8-, 12-, or 96-tip liquid-handling robotic devices (Figure 1B)

    Both E-PAGE™ 48 and 96 cassettes are conveniently opened with a butterfly opener to remove the gel for downstream staining or blotting (Figures 4 and 5). In addition, each cassette is labeled with an individual barcode to facilitate identification of the gel using commercial barcode readers.

Table 1 - Specifcations of the E-PAGE™ 48 8% and E-PAGE™ 96 6% Gels


 E-PAGE 48 8% Gels E-PAGE 96 6%Gels
Cassette size 13.5 (l) x 10.8 (h) x 0.67 cm (w) 13.5 (l) x 10.8 (h) x 0.67 cm (w)
Gel thickness 3.7 mm 3.7 mm
Gel volume 50 ml 50 ml
Well depth 3 mm 3 mm
Number of wells 48 + 4 markers 96 + 8 markers
Well format 2 x 26 Staggered 8 x 13
Well opening 3.6 mm x 2.2 mm 3.8 mm x 1.8 mm
Running distance 32 mm 16 mm
Run time 23 minutes 14 minutes
Space between well centers 4.5 mm 9 mm
Sample volume 15 µl 20 µl
Max protein load 20 µg 20 µg

  • may require an orientation adaptor
Table 2 - Migration patterns of protein standards on the E-PAGE™ 48 8% and E-PAGE™ 96 6% Gels

E-Base™ Integrated Devices

  • E-PAGE™ Gels run in the specially designed E-Base™ electrophoresis bases, which combine the base and the power source in one device. The E-Base™ devices are compatible with the Society for Biomolecules Screening (SBS)-standard plate size and can be conveniently mounted on liquid handling robot decks. Two types of bases are available from Invitrogen:
    1. The mother E-Base™ device contains an electrical plug that can be connected directly to an electrical outlet and is used for electrophoresis of one E-PAGE™ Gel
    2. The daughter E-Base™ device connects to the mother E-Base™ (Figure 3) and to other daughter E-Base™ devices for the simultaneous electrophoresis of two or more E-PAGE™ Gels. The daughter E-Base™ does not contain an electrical plug and cannot be used without a mother base

    Each base has a power/program button and a timer button on the lower right side of the base for controlling your electrophoresis run. The lower left side of each base contains a lighted LED and a digital timer display (00-99) that indicates where your gel is in the running process. Two programs are pre-set in the E-Base™ units: a 14-minute protein run program for E-PAGE™ 96 gels and a 12-minute DNA program for running E-Gel® 96 Gels. For E-PAGE™ 48 Gels, use the protein program extended to 23 minutes due to the longer running distance.

    Note: The E-Base™ can run E-Gel® 96, E-Gel® 48, E-PAGE™ 96, or E-PAGE™ 48 systems.
    The E-PAGE™ gels are not compatible with old models of the E-Gel® 96 bases (mother and daughter bases).

E-Holder™platform

  • The E-Holder™ platform is compatible in size with standard automated systems for convenient loading of samples in a hands-free robot environment when the use of the E-Base™ devices is not practical or while in queue for its availability.

E-Editor™ 2.0 Software

  • Analysis of E-PAGE™ gel results is fast and convenient, for both stained gels and blots, using the Window®-based E-Editor™ 2.0 software. All lanes, including sample lanes, are grouped and aligned into one image for easy interpretation and comparison. Moreover, when 384-well assays are performed, the E-Editor™ program allows selection of multiple automated loading patterns and grouping of four different E-PAGE™ 96 Gel results or eight different E-PAGE™ 48 Gel results into one integrated image. This enables an immediate reference back from the gel or blot lane into the coordinates of the originating plate well to provide quick identifcation of important results.
Important Licensing Information
These products may be covered by one or more Limited Use Label Licenses (See Catalog Number/Label License Index and Label Licenses in Appendix). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. All products are for research use only. CAUTION: Not intended for human or animal diagnostic or therapeutic use.

Figure 1. Loading E-PAGE™ Gels using a multichannel pipettor or liquid-handling robot

Figure 2. The staggered-well format of an E-PAGE™ 96 pre-cast gel

  • 10 µ E-PAGE™ SeeBlue® Pre-Stained Standard was loaded into some wells of an E-PAGE™ 96 6% Gel.

Figure 3. The mother/daughter E-Base™ combination

Figure 4. Protein staining results on E-PAGE™ Gels

  • A. Expressway™ HTP Mix was used to express various human kinase ORFs with the pEXP3-DEST Expressway™ vector. Reactions were electrophoresed on an E-PAGE™ 96 6% gel and analyzed using the Lumio™ Green Detection Kit.
    B.
    		 Lanes M: 5 µl SeeBlue® Plus2 Pre-stained Std.
    Lanes 2, 4, 21, 23: 10 µl MagicMark™ XP Std.
    Lanes 6 & 19: 2 µl BenchMark™ His-tagged Std.
    Lane 8: lysozyme 200 ng
    Lane 10: Carbonic Anhydrase 200 ng
    Lane 12: BSA 100 ng
    Lanes 15 & 17: 5 µl of E. coli lysate Other lanes: blank The gel stained with Commassie® R-250 following the recommended protocol in the E-PAGE™ 48 manual.

Figure 5. Western blot results on E-PAGE™ Gels

  • 5 µl MagicMark™ XP Western Standard was loaded into each well of an E-PAGE™ 48 8% Gel and transferred to a pre-cut 0.2 µm nitrocellulose membrane (Cat. no. LC2009). The blot was probed with 1:1000 Anti-Xpress™ Antibody and detected using the WesternBreeze® Immunode-tection Kit (anti-mouse) – Chemiluminescent (A) and Chromogenic (B) detection.