Abstract: An apparatus and method are described for capillary separation of macromolecules and precise post-separation blotting. Apparatus include disposable separating element (capillary), which contains a sieving or interaction matrix inside, an external layer of blotting material, positioned close to the boundary of said sieving or interaction matrix, and the membrane with changeable permeability for separated material; said membrane separates blotting layer from the sieving or interaction matrix. After separation of macromolecules in capillary with initially non-permeable walls, chemical or physical modification of the membrane is performed, which is followed by changing the vector of driving forces for transfer, so that separated molecules are moved through the walls of the capillary and blotted to the outer layer of separating element, which contains blotting material. Means of modification of the membrane include chemical or physical modification, leading to changes in permeability.

Abstract: The present invention discloses a semi-dry electroblotter for transferring a gel after its electrophoresis onto a membrane enabling the electro-blotting process to be efficiently and effectively proceeded within a short time, and eliminating the shortcomings of the traditional technology. The electroblotter of the present invention comprises an upper block, and a corresponding lower block in the shape of a groove. A corresponding positioning member is disposed on each of the lateral sides of the upper block and the lower block, such that the upper block and the lower block will be coupled automatically when they are closed in a correct guiding position. An elastic member and an electrode plate are separately disposed in the interior of the upper block and the lower block, and the elastic member supports the electrode plate so that the electrode plate has an elastic reaction on the opposite direction when the electrode plate is pressed.

Abstract: A multi-dimensional proteomic analysis method utilizing cationic electrophoresis is described. The method includes separating proteins in one direction using cationic electrophoresis and separating the proteins in a second orthogonal direction using other electrophoresis separation methods such as denaturing electrophoresis and electrophoresis subsequent to proteolytic cleavage or isofocussing. The two dimensional array may be used to determine various protein-protein interactions in a sample.

Abstract: An automated high-throughput system for excising spots or samples from an electrophoresis slab gel includes a computer controlled robotic arm assembly and a sample plate handling assembly for supplying a sample plate to a loading station. The computer is connected to a scanner and imaging device to identify selected sample locations on the slab gel and to direct the robotic arm to the selected locations for excising the gel spots. The cutting assembly includes a removable tray for supporting the slab gel during the cutting process and is coupled to the automated sample plate handling assembly. The sample plate handling assembly delivers a multiwell plate to the cutting assembly for receiving the gel spots. The removable tray cooperates with a scanner for identifying protein spots and includes a positioning device to position the tray in the scanner and the cutting assembly in selected locations to coordinate the scanned image with the cutting process.

Abstract: A gel relaxer device is provided for relieving stress in an electrophoresis gel between clamping surfaces of a gel clamp. The device includes a pair of operating arms having a gel gripping surface and a clamping actuating surface. The operating arms move toward each other to enable the gripping surfaces to grip and support an electrophoresis gel suspended by a gel clamp. The actuating surfaces of the arms engage the clamping jaws of the gel clamp to open the jaws to allow the gel to relax and assume its normal position and orientation. The operating arms are then separated to allow the gel clamp to again close and clamp the gel. The gel and gel clamp can then be transported to a selected processing stage.

Abstract: The present invention provides an integrated, fully automated, high-throughput system for two-dimensional electrophoresis comprised of gel-making machines, gel processing machines, gel compositions and geometries, gel handling systems, sample preparation systems, software and methods. The system is capable of continuous operation at high-throughput to allow construction of large quantitative data sets.

Abstract: The present invention provides an integrated, fully automated, high-throughput system for two-dimensional electrophoresis comprised of gel-making machines, gel processing machines, gel compositions and geometries, gel handling systems, sample preparation systems, software and methods. The system is capable of continuous operation at high-throughput to allow construction of large quantitative data sets.

Abstract: The present invention provides an integrated, fully automated, high-throughput system for two-dimensional electrophoresis comprised of gel-making machines, gel processing machines, gel compositions and geometries, gel handling systems, sample preparation systems, software and methods. The system is capable of continuous operation at high-throughput to allow construction of large quantitative data sets.

Abstract: A method for drying a polyacrylamide gel, the method comprising contacting the gel with an aqueous solution of a polyhydoxy alcohol other than a polyhydroxy alcohol having at least 3 vicinal hydroxy groups and drying the gel. Examples of suitable polyhydroxy alcohols include 1,2-ethane diol, 1,2-propane diol, 1,3-propane diol, 1,4-butane diol, 1,6 hexane diol, alkyl triol, 1,2,6-trihydroxy hexane, trimethylol propane and pentaerythritol. The gel may be dried between two cellophane sheets positioned in a drying frame arrangement.

Abstract: The invention relates to a sample retrieval apparatus of particular benefit in the field of molecular biology. The apparatus permits the rapid collection of numerous specified fractions of samples: such as DNA, RNA or protein, that are separated by gel electrophoresis; or microorganisms grown on agar plates. The apparatus is capable of multiple sample retrieval from gels without cross-sample contamination from previously excised samples. Specifically, the sample retrieval apparatus is able to engage a cutting tip, cut a desired spot, band or plaque from a gel, deposit the desired band or plaque into a container, such as a multi-well plate for processing, and disengage the used cutting tip. The apparatus is then able to repeat this process many times to facilitate rapid and accurate processing of multiple bands or plaques from a single gel using different cutting tips for each sample retrieval.

Abstract: A gel process plate is formed of a base member with a plurality of concave portions, and a lid member with a plurality of convex portions. Each concave portion has a bottom surface provided with holes for allowing a liquid to pass therethrough. Each convex portion has a height less than a depth of the concave portion of the base member, and a top surface with holes for allowing the liquid to pass therethrough. The convex portions are disposed at portions corresponding to the concave portions of the base member to fit the concave portions. When the base member and the lid member are assembled, a space is defined between the bottom surface of the concave portion and the top surface of the convex portion to hold a piece, i.e. gel piece, therein for processing.

Abstract: The present invention relates to methods for separating nucleic acids from other cellular debris, especially substances that carry a net positive charge at low pH, by electrophoresis under acid conditions. In the purification method of the present invention, nucleic acids are separated from proteins found in the same biological sample by applying the sample to an electrophesis gel and subjecting the sample to electrophoresis under acid conditions to separate the nucleic acids from the proteins. The optimum pH may differ for different sample types but can be readily determined by those skilled in the art. Preferably, the separation is performed at a pH of about 2 to about 4. More preferably, electrophoresis is carried out at a pH of 2.

Abstract: The present invention provides an integrated, fully automated, high-throughput system for two-dimensional electrophoresis comprised of gel-making machines, gel processing machines, gel compositions and geometries, gel handling systems, sample preparation systems, software and methods. The system is capable of continuous operation at high-throughput to allow construction of large quantitative data sets.

Abstract: A method and device for transferring an electrophoresis gel from a cassette to a carrier includes a tray having a dimension to receive a cassette and a carrier device. The tray has a bottom wall with a surface with channels or recesses formed by projections to reduce contact of an electrophoresis gel with the bottom wall of the tray. The projections are typically pyramid-shaped members extending to a peak having a small surface area to prevent the electrophoresis gel from adhering to the bottom wall. The bottom wall also includes a recessed area to receive the carrier so that the gel can slide easily from the cassette to open jaws of the carrier. A retaining arm extends from a side wall of the tray to hold the jaws of the carrier in an open position while the gel is being positioned between the clamping surfaces of the jaws.

Abstract: The present invention provides an integrated, fully automated, high-throughput system for two-dimensional electrophoresis comprised of gel-making machines, gel processing machines, gel compositions and geometries, gel handling systems, sample preparation systems, software and methods. The system is capable of continuous operation at high-throughput to allow construction of large quantitative data sets.

Abstract: The present invention provides an integrated, fully automated, high-throughput system for two-dimensional electrophoresis comprised of gel-making machines, gel processing machines, gel compositions and geometries, gel handling systems, sample preparation systems, software and methods. The system is capable of continuous operation at high-throughput to allow construction of large quantitative data sets.

Abstract: The present invention provides electrophoresis apparatus and electrophoresis methods employing gellan gum based gels employing divalent metal cation and diamine cross-linking agents. The gels are reversible under conditions that do not damage the biomolecules separated using the gels. The present invention also provides novel gellan gum-based gels which are cross-linked which employ a diamine cross-linking agent.

Abstract: An assay for the identification of neutralizing botulinum antibodies in sera is provided which includes the steps of separating non-sodium dodecyl sulfate, non-trypsinized complex botulinum toxin in an arylamide gel by electrophoresis, the separation occurring on a basis of botulinum toxin protein size and charge. Thereafter, the separated protein is electrophoretically transferred onto a solid support, the transferred separated protein being bound to the solid support at spaced apart sites. Remaining sites on the solid support not occupied by bound protein are blocked and the solid support and bound protein are contacted with a sera sample containing an antibody directed against botulinum toxin. The contacted solid support is then exposed to a second antibody capable of reacting to produce an insoluble colored substrate of intensity relative to a quantity of antibodies present. Finally, the second antibody is reacted to produce the insoluble colored substrate which is visually identified.

Abstract: The present invention provides an integrated, fully automated, high-throughput system for two-dimensional electrophoresis comprised of gel-making machines, gel processing machines, gel compositions and geometries, gel handling systems, sample preparation systems, software and methods. The system is capable of continuous operation at high-throughput to allow construction of large quantitative data sets.

Abstract: The claimed multi-tank gel developing apparatus and method comprises of two inner tanks within one outer tank to develop multiple electrophoresis gels simultaneously. The two inner tanks have openings along the edges of the walls to allow free circulation of solutions between the inner and outer tanks. The solution maybe exchanged by lifting out the inner tanks which retains the gels inside while the solution drains out. Once the solution in the outer tank has been exchanged, the inner tanks can be re-immersed into the outer tank. Furthermore, the inner tanks have detachable bases to facilitate transfer of the gels after development. This claimed apparatus allows for efficient development of electrophoresis gels by minimizing the direct handling of the gels thereby avoiding unwanted damages often incurred by conventional manually developing methods and apparatuses. The multi-tank apparatus can accommodate various gel compositions and sizes.

Abstract: The present invention presents a novel method for both staining electrophoresis gels, as well as removing the background dye during de-staining. This novel and simple pad-based technique minimizes solvent consumption, is environmentally friendly, and does not leave any solid residues on the gel. Furthermore, a novel method for the quantitative and qualitative determination of protein concentration in solution is also presented. This method is simple, easy-to-use and has an easily visible color change from clear to blue in the presence of protein.

Abstract: The present invention provides electrophoresis systems and methods for forming at least one gel therein and for conducting electrophoresis on the gel without removing the gel from the electrophoresis system. The systems of the present invention include an electrophoresis platform which is a surface onto which one or more electrophoresis gels can be cast, and at least one gel casting/electrophoresis member for use in casting at least one gel on the electrophoresis platform and conducting electrophoresis on the gel. The present invention also includes optional members specific to other stages of the electrophoresis process, including a gel staining member and a gel drying member.

Abstract: A process for electroelution of a gel containing charged macromolecules, such as proteins or polynucleotides, comprising the steps of providing a plurality of adjacent parallel chambers having a trapezoidal cross-section, placing a gel containing the charged macromolecules onto first open sides of the chambers, placing a semipermeable membrane onto second open sides of the chambers, filling the chambers with an elution buffer and applying a voltage difference across the chambers so that charged macromolecules in the gel migrate into the elution buffers in the chambers. Also, an apparatus for electroelution of a gel containing charged macromolecules having, preferably, a plurality of adjacent parallel chambers having a trapezoidal cross-section and vents for removing the product without disassembling the apparatus.

Abstract: A new process and kit are described that combines methods for generating the nucleotide base sequence of a DNA molecule with an ultra-sensitive silver staining protocol. This new combination of technologies allows for a direct, non-instrument based visualization of electrophoretically separated sequencing fragments. This non-radioactive system includes sequencing the DNA molecule by forming a set of fragments using an enzymatic dideoxy-mediated chain termination method, electrophoretically separating the DNA fragments on a gel medium, and exposing the gel medium to ultra-sensitive silver-staining solutions for a time determined by viewing the silver stain reacted primer extension products.

Abstract: An apparatus for electroelution isolation of biomolecules and recovering biomolecules after elution including a tubular enclosure for engaging a piece of separating gel having a band of biomolecules. The enclosure is capped by a closure means having a passage means, and then placed in an electrophoresis tank with the closure means facing the positive terminal. Applying an electric current forces the biomolecules to accumulate at the closure means. The biomolecules may be recovered by inserting a capillary tip through the passage means and drawing the biomolecules through the capillary.

Abstract: A polyacrylamide gel which has undergone electrophoresis is dried in the presence of pressure applied via a semipermeable film placed on the gel. Before applying the semipermeable film onto the gel, solution components contained in the gel is replaced by an aqueous solution containing one or more substances selected from the group consisting of saccharides, sugar alcohols having 4 or more carbon atoms, and water-soluble polymers. According to this method, a polyacrylamide gel which has undergone electrophoresis can be dried with ease and reduced costs while maintaining its transparency and without causing cracks even in the case of a gel containing polyacrylamide gel at a high concentration.

Abstract: The invention relates to the mass spectrometric analysis of separated substance samples on so-called 2-D-gel electrophoresis plates, used particularly for protein determination, with ionization of the substance samples by MALDI. The molecules of the substance samples are transferred to a thin, lacquer-like smooth matrix layer which should preferably be applied to a metal sample support. Transfer takes place directly or indirectly, for example via a polyvinylidene difluoride membrane an an intermediate carrier, by electrophoretic transport of the molecules to the matrix surface. Before transfer, the proteins may be subjected to enzymatic cutting of their amino acid chains.

Abstract: A multiple electrophoresis method and apparatus for migrating and transferring macromolecules in a vessel (10) containing a plurality of parallel elongate electrodes (14), membranes (22) disposed vertically in the vessel between the columns of electrodes (14), and an electrophoresis gel which is inserted in the vessel in liquid form and which is subsequently solidified to perform electrophoresis. When the electrophoresis is completed, the gel is liquefied, dissolved, or decomposed, and the membranes (22) onto which the macromolecules have been transferred are withdrawn from the vessel.

Abstract: An apparatus for collecting samples by gel electrophoresis has a base table, a cutting tool or an extractor disposed above the base table, a moving mechanism, a pattern detector and a control device. The base table is for placing thereon a gel in the form of a slab with one of its sandwiching support plates removed therefrom in such a way that the gel is exposed in the upward direction. The cutter is for cutting out a specified portion of the gel, and the extractor is for extracting a specified migration band in such a specified portion of the gel by electrophoresis. The moving mechanism is for moving the cutter, or the extractor, three-dimensionally. The pattern detector is for optically detecting migration patterns in the gel on the base table so as to enable the user to determine which part of the gel or which migration band thereon is of interest.

Abstract: A novel device and method for biomolecule purification is provided as a handy tool for routine applications when both a high resolution and an easy manipulation are required. A sample is applied into a device and the different biomolecules are separated from each other and stored in segregated liquid fractions in the device automatically after 1 hour electrophoresis. These purified biomolecules in liquid fractions are then easily released from the device in 4 minutes.

Abstract: Separation media for electrophoresis, and methods of filling and flushing of electrophoretic devices such as capillaries are described. By preparing submicron to above-micron sized cross-linked gel particles and using gel swelling equilibrium concepts, such devices can be easily filled and flushed. Gel particles can be prepared by inverse suspension, precipitation and suspension polymerization. These particles can be swollen and collapsed by small changes in temperature, pH, and ionic strength of solvent. Other approaches involve the formation of reversible cross-links by use of polyelectrolyte complexes, chelating agents or copolymers of hydrophobic and hydrophilic repeat units. Finally, reversibly solubilized systems may be used to change the viscosity of the media.

Abstract: A silver staining method is provided for staining an organic molecule capable of binding silver. An improved image is developed and visualized. A kit useful in practicing the method is described. A permanent record of the image of the profile of the stained molecules is obtained.

Abstract: A method for covalent immobilization of a carbohydrate molecule with an oppositely charged surface, comprising the steps of adsorbing the carbohydrate molecule to the oppositely charged surface in proximity to reactive moieties bound to the charged surface; and next activating the bound moiety sufficiently for covalent attachment to the adsorbed carbohydrate molecule. Also disclosed is a hydrophobic microporous polymer membrane coated with a cross-linked, cationic polymer having fixed charges thereon, the coating having enhanced epoxide content for covalent immobilization of the oppositely-charged carbohydrate molecule. A novel method for sequencing carbohydrates by covalent attachment of a carbohydrate conjugate to the membrane, and subsequent treatment with glycosidases, is also presented.

Abstract: An elution cassette is invented for macromolecule recovery from electrophoresis gels. The cassette comprises a frame (10), a front barrier (20), a back barrier (30), and a space inside the cassette (35) defined by the frame and the barriers. The barrier made of filter materials allows macromolecules to pass through while stopping gel debris. The back barrier is made of a membrane material allowing small ions to pass through but not macromolecules. When the cassette is inserted into an agarose gel and the electrophoresis is continued, macromolecules driven by an electric field migrate into the cassette through the front barrier and are stopped by the back barrier. The macromolecules contained in the space between the barriers can then be recovered.

Abstract: A preparative two dimensional gel electrophoresis system which serves as a single procedure for separation and isolation of preparative amounts of proteins from complex biological preparations. The system includes sized-up isoelectric focusing tube gels and slab gel molds which allow for sample loads of between about 0.5 and 2 mg or greater. Increased protein loads, resolution and electrotransfer allow for subsequent sequencing of separated proteins by conventional methods.

Abstract: A flow-through receptacle is disclosed, one end of which is designed to hold multiple plugs of gel material, containing solutes such as macromolecules, excised from electrophoretic separations, and the other end of which is capable of insertion into a recipient matrix that lies between the plates of a slab gel enclosure in an electrophoresis apparatus. The receptacle is used to place the gel plugs, containing the macromolecules or solutes, into electrophoretic contact with a recipient matrix and allows for electrophoretic transfer of the solutes from the plugs into a single concentrated zone in the recipient matrix. The concentrated macromolecules or solutes can then be further processed in the recipient gel or excised and used for procedures requiring greater amounts and concentrations of the solute than are available in the original plugs.

Abstract: A new process and kit are described that combines methods for generating the nucleotide base sequence of a DNA molecule with an ultra-sensitive silver staining protocol. This new combination of technologies allows for a direct, non-instrument based visualization of electrophoretically separated sequencing fragments. This non-radioactive system includes sequencing the DNA molecule by forming a set of fragments using an enzymatic dideoxy-mediated chain termination method, electrophoretically separating the DNA fragments on a gel medium, and exposing the gel medium to ultra-sensitive silver-staining solutions for a time determined by viewing the silver stain reacted primer extension products.

Abstract: An electrophoresis apparatus for automatically performing medical assays includes an electrophoresis platform which cooperates with a gantry assembly. The electrophoresis platform and the gantry assembly are movable along paths that are perpendicular to each other. An applicator assembly includes pipettes which transfer fluid samples from a specimen tray to an electrophoresis plate mounted on the electrophoresis platform. The electrophoresis platform then moves to a position into the gantry assembly, where electrophoresis is conducted to separate the samples into different fractions. The electrophoresis platform then moves beneath a reagent pouring station where a reagent is applied to make the separated fractions fluoresce under ultraviolet light. The electrophoresis platform is then moved beneath the gantry assembly again, and an air knife in the gantry assembly spreads the reagent.

Abstract: A method for visualizing nucleic acids in a polyacrylamide gel. The method comprises fixing the nucleic acids with 10% acetic acid for about 20 minutes, washing the gel multiple times with water for about 2 minutes, impregnating the gel with silver nitrate at a concentration of about 1.0 g/l and about 1.5 ml/l of 37% formaldehyde for about 30 minutes, developing the gel with sodium carbonate at a concentration of about 30 g/l, 1.5 ml/l of 37% formaldehyde and sodium thiosulfate pentahydrate at a concentration of about 2.0 mg/l for between about 2 minutes and about 5 minutes, and then stopping the development of the gel by treatment with 10% acetic acid for about 5 minutes.