Bio-rad Laboratories, Inc.

James F Monthony

Timothy E Delony

Robert V Dahlstrom

Michael W Burgett

Christopher J Siebert

Nathan Lewin

Paul F Wegfahrt

Eric G Wallace

Donald F Hollis

James R Gangwer

Gian D Polastri

Walter H Schick

Norman D Schwartz

An assay method suitable for antigens (or haptens) including proteins and polypeptides utilizing antibodies for the unknown protein or polypeptide in which the antibodies are covalently bound to water insoluble hydrophilic polymeric particles. Appropriate fluorescently labeled immune reactants are introduced during the method and together with the unknown antigen are immunologically bound, directly or indirectly, to the particles for separation from the reaction liquid. The particles have a size of about 0.1-10 microns and can be resuspended for direct measurement of the fluorescent labels in a fluorometer from which the unknown can be determined.

Reagent combination for solid phase immunofluorescent assay

A method and apparatus for casting gel slabs useful in electrophoretic procedures is provided. Glass plates are horizontally positioned in parallel a capillary action-inducing distance apart to form an unsealed space. Gel-forming liquid, e.g. polyacrylamide is introduced between the plates and fills the space by capillary action. The surface tension of the gel-forming liquid is utilized to constrain the liquid between the plates at the unsealed or open edges. Once the desired sized space is filled the gel is formed by polymerization and/or cooling. Where an open faced gel slab is desired one of the plates is separated from the gel layer.

Gel slab casting

Insoluble cationic acrylamide copolymers for use as cell growth carriers and a novel copolymer of dimethylaminopropylmethacrylamide, acrylamide and methylene-bis-acrylamide useful as both a carrier for cell growth and as a medium for ion exchange chromatography of aqueous protein solutions.

Micro-carrier cell culture

An assay method suitable for antigens (or haptens) including proteins and polypeptides utilizing antibodies for the unknown protein or polypeptide in which the antibodies are covalently bound to water insoluble hydrophylic polymeric particles. Appropriate fluorescently labeled immune reactants are introduced during the method and together with the unknown antigen are immunologically bound, directly or indirectly, to the particles for separation from the reaction liquid. The particles have a size of about 0.1-10 microns and can be resuspended for direct measurement of the fluorescent labels in a fluorometer from which the unknown can be determined.

Solid phase immunofluorescent assay method

Electrophoresis apparatus having a housing, a substantially horizontal cooling bed within the housing for supporting and cooling a medium during electrophoresis, a transparent cover overlying the bed, and first and second electrode troughs along opposed sides of the bed. A condenser coil has an open central area greater than the area of the bed and overlies the bed with the open area located above a vertical projection of the bed. Major segments of the condenser coil overlie the electrode troughs. The condenser coil and the cooling bed are arranged for serial fluid flow with coolant fluid introduced into the condenser coil and withdrawn from the cooling bed. The cooling bed itself has first and second generally parallel plates spaced apart by a core which defines a tortuous path for fluid flow. One of the plates has a higher thermal conductivity than the other. By inverting the cooling bed, differing cooling rates can be achieved. Electrode wires within the electrode troughs are connected at diagonally opposite ends to a DC power supply to eliminate nonuniformities in the electric potential from one end of the medium to the other.

Electrophoresis apparatus

The method of this invention is characterized by the use of hydrolyzed cross-linked polyacrylamide particles to which have been bonded, by means of covalent bonds, antibiodies against the thyroid hormone to be determined. The particles selected are of a size which forms a stable hydrophyllic suspension. In the preferred embodiment a measured quantity of unextracted human serum is mixed together with a blocking agent in an amount sufficient to displace the thyroid hormone to be measured from thyroxine binding globulin (TBG), a radioactively labeled thyroid hormone of the type to be measured, and the antibody-polyacrylamide complex. The hormone to be measured is displaced by the blocking agent followed by the competitive binding of the labeled and unlabeled hormone to the antibody-polyacrylamide particles. The particles are readily separated from the sample liquid and the radioactivity of the particle material and/or in the liquid is determined. Particles containing Alcian yellow or blue dye facilitate practice of the method.

Radioimmunoassay for thyroid hormone

Electrophoretic gels are destained between two layers of ion exchange resin. Electrical current is passed from one layer of resin to the other through the gel in an electrically low-conductive surrounding medium such as de-ionized water. Virtually all of the current goes through the gel resulting in the highly efficient destaining technique.

Method for electrophoretic destaining

Gels for electrophoresis such as polyacrylamide gels are formed in a neutral pH medium containing a salt solution of preselected concentration. This salt is incorporated in the gel and in subsequent use in electrophoresis is displaced upon application of an electrical potential (direct current) by ionic species which form a pH buffer in the gel. Prior to introduction of the buffer, the gel containing the neutral salt is storage stable for prolonged periods of time. The gel is reproducible since it is always formed under the same conditions and the buffer only thereafter introduced.

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Albany, CA, United States of America

James F Monthony