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US 6,635,489 B2Granted
Materials for enhancing staining of biopolymers in matrices
AssigneeInvitrogen Corporation
InventorScott Whitney
Life of this patent
Estimating the term…
Abstract
Methods for detecting biopolymers in a matrix are disclosed, which involve contacting the matrix with a sensitizing reagent, which may include one or more optionally substituted heteroaromatic compounds; contacting the matrix with one or more reduceable metal salts to stain the biopolymer; and detecting the stained biopolymer. Also disclosed are compositions for carrying out the invention and compositions made according to the invention. Also disclosed are kits for carrying out the methods of the invention.
Claims51
A method for detecting a biopolymer in a matrix, comprising:
- (a) contacting the matrix with a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) contacting the matrix with one or more reduceable metal salts to stain said biopolymer;
- and
- (c) detecting the stained biopolymer.
1 more element
A method for detecting a protein/peptide or nucleic acid molecule in a polyacrylamide gel, comprising:
- (a) fixing the protein/peptide or nucleic acid molecule to the polyacrylamide gel;
- (b) contacting the polyacrylamide gel with a sensitizing reagent comprising primuline, thioflavin S or 2-(4-aminophenyl)-6-methyl-7-sulfobenzothiazole;
- (c) contacting the polyacrylamide gel with an aqueous solution of a silver salt to stain the protein/peptide or nucleic acid molecule,
- (d) developing the stained protein/peptide or nucleic acid molecule, and
- (e) detecting the stained protein/peptide or nucleic acid molecule.
2 more elements
A method for identifying a protein or peptide in a matrix, comprising:
- (a) contacting the matrix with a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) contacting the matrix with one or more reduceable metal salts to stain said protein or peptide;
- (c) detecting the stained protein or peptide;
- (d) carrying out a cleavage reaction on the protein or peptide to give fragments;
- and
- (e) carrying out a mass spectrometric analysis on said fragments thereby identifying the protein or peptide.
3 more elements
A composition, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more developer solutions comprising a reducing agent;
- (d) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- (e) one or more contrast enhancing agents;
- (f) one or more buffers;
- (g) one or more fixing reagents;
- (i) one or more biopolymers;
- (j) one or more matrixes;
- and
- (k) phenolphthalein or thymolphthalein.
8 more elements
A composition, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more developer solutions comprising a reducing agent;
- (d) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- (e) one or more contrast enhancing agents;
- (f) one or more buffers;
- (g) a fixing reagent comprising an aqueous solution of an organic acid and a lower alcohol containing 1-4 carbon atoms;
- (i) one or more biopolymers;
- (j) one or more matrixes;
- and
- (k) one or more indicators which are sensitive to pH changes.
8 more elements
A composition, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more developer solutions comprising a reducing agent;
- (d) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- (e) one or more contrast enhancing agents;
- (f) one or more buffers;
- (g) a fixing reagent consisting of about 40% ethanol, about 10% acetic acid and about 50% distilled water by volume;
- (i) one or more biopolymers;
- (j) one or more matrixes;
- and
- (k) one or more indicators which are sensitive to pH changes.
8 more elements
A composition, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds, wherein said optionally substituted heteroaromatic compounds are primuline, thioflavin S or 2-(4-aminophenyl)-6-methyl-7-sulfobenzothiazole;
- (b) one or more reduceable metal salts;
- and
- (c) one of more developer solutions comprising a reducing agent.
1 more element
A composition, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds wherein said optionally substituted heteroaromatic compounds are primuline, thioflavin S or 2-(4-aminophenyl)-6-methyl-7-sulfobenzothiazole;
- (b) one or more developer solutions comprising a reducing agent;
- and
- (c) one or more contrast enhancing agents selected from the group consisting of sodium sulfide, thiourea, dithiothreitol, potassium tetrathionate, sodium dithionite, and the sodium or potassium salt of thiosulfate.
1 more element
A composition, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- and
- (d) one or more contrast enhancing agents selected from the group consisting of sodium sulfide, thiourea, dithiothreitol, potassium tetrathionate, sodium dithionite, and the sodium or potassium salt of thiosulfate.
2 more elements
A composition, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more developer solutions comprising a reducing agent;
- (d) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- (e) one or more contrast enhancing agents selected from the group consisting of sodium sulfide, thiourea, dithiothreitol, potassium tetrathionate, sodium dithionite, and the sodium or potassium salt of thiosulfate;
- (f) one or more buffers;
- (g) one or more fixing reagents;
- (i) one or more biopolymers;
- (j) one or more matrixes;
- and
- (k) one or more indicators which are sensitive to pH changes.
8 more elements
A composition, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more developer solutions comprising a reducing agent;
- (d) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- (e) one or more contrast enhancing agents;
- (f) one or more buffers having a pKa of 5-10;
- (g) one or more fixing reagents;
- (i) one or more biopolymers;
- (j) one or more matrixes;
- and
- (k) one or more indicators which are sensitive to pH changes.
8 more elements
A composition, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more developer solutions comprising a reducing agent;
- (d) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- (e) one or more contrast enhancing agents;
- (f) one or more buffers selected from the group consisting of morpholinoethanesulfonic acid, 4-(2-hydroxyethyl)-1-piperazinepropane-sulfonic acid (EPPS) and 4-(2-hydroxy-ethyl)-1-piperazineethanesulfonic acid (HEPES);
- (g) one or more fixing reagents;
- (i) one or more biopolymers;
- (j) one or more matrixes;
- and
- (k) one or more indicators which are sensitive to pH changes.
8 more elements
A composition, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- and
- (d) one or more contrast enhancing agents.
2 more elements
A kit for the detection of biopolymers, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more developer solutions comprising a reducing agent;
- (d) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- (e) one or more contrast enhancing agents;
- (f) one or more buffers;
- (g) one or more fixing reagents;
- (h) one or more cleaving reagents;
- (i) one or more biopolymers;
- (j) one or more matrixes;
- and
- (k) phenolphthalein or thymolphthalein.
9 more elements
A kit for the detection of biopolymers, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more developer solutions comprising a reducing agent;
- (d) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- (e) one or more contrast enhancing agents;
- (f) one or more buffers;
- (g) a fixing reagent comprising an aqueous solution of an organic acid and a lower alcohol containing 1-4 carbon atoms;
- (h) one or more cleaving reagents;
- (i) one or more biopolymers;
- (j) one or more matrixes;
- and
- (k) one or more indicators which are sensitive to pH changes.
9 more elements
A kit for the detection of biopolymers, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more developer solutions comprising a reducing agent;
- (d) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- (e) one or more contrast enhancing agents;
- (f) one or more buffers;
- (g) a fixing reagent consisting of about 40% ethanol, about 10% acetic acid and about 50% distilled water by volume;
- (h) one or more cleaving reagents;
- (i) one or more biopolymers;
- (j) one or more matrixes;
- and
- (k) one or more indicators which are sensitive to pH changes.
9 more elements
A kit for the detection of biopolymers, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more developer solutions comprising a reducing agent;
- (d) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- (e) one or more contrast enhancing agents selected from the group consisting of sodium sulfide, thiourea, dithiothreitol, potassium tetrathionate, sodium dithionite, and the sodium or potassium salt of thiosulfate;
- (f) one or more buffers;
- (g) one or more fixing reagents;
- (h) one or more cleaving reagents;
- (i) one or more biopolymers;
- (j) one or more matrixes;
- and
- (k) one or more indicators which are sensitive to pH changes.
9 more elements
A kit for the detection of biopolymers, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more developer solutions comprising a reducing agent;
- (d) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- (e) one or more contrast enhancing agents;
- (f) one or more buffers having a pKa of 5-10;
- (g) one or more fixing reagents;
- (h) one or more cleaving reagents;
- (i) one or more biopolymers;
- (j) one or more matrixes;
- and
- (k) one or more indicators which are sensitive to pH changes.
9 more elements
A kit for the detection of biopolymers, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds;
- (b) one or more reduceable metal salts;
- (c) one or more developer solutions comprising a reducing agent;
- (d) one or more stopper solutions which prevent further reduction of the reduceable metal salts;
- (e) one or more contrast enhancing agents;
- (f) one or more buffers selected from the group consisting of morpholinoethanesulfonic acid, 4-(2-hydroxyethyl)-1-piperazinepropane-sulfonic acid (EPPS) and 4-(2-hydroxy-ethyl)-1-piperazineethanesulfonic acid (HEPES);
- (g) one or more fixing reagents;
- (h) one or more cleaving reagents;
- (i) one or more biopolymers;
- (j) one or more matrixes;
- and
- (k) one or more indicators which are sensitive to pH changes.
9 more elements
A kit for the detection of biopolymers, comprising:
- (a) a sensitizing reagent comprising one or more optionally substituted heteroaromatic compounds wherein said optionally substituted heteroaromatic compounds are primuline, thioflavin S or 2-(4-aminophenyl)-6-methyl-7-sulfobenzothiazole;
- and
- (b) silver nitrate.
Dependencies are inferred from the claim text.