CN104483479A - Dot immunogold filtration assay method for testing antique silk fabric - Google Patents

Abstract

The invention discloses a dot immunogold filtration assay method for testing antique silk fabric. A micro-pore filtration membrane serves as a carrier, antigens or antibodies are dotted on the membrane, a to-be-tested sample is additionally arranged after the membrane is sealed, and the corresponding antigen is detected with the antibody marked with colloidal gold after washing; qualitative analysis can be conducted according to existence of red color on the carrier. The dot immunogold filtration assay method for testing the antique silk fabric has the beneficial effects of high sensitivity, simplicity of operation, low cost and short reaction time.

Description

A kind of dot immuno gold filtration assay measures the method for Ancient Silk Textile

Technical field

The invention belongs to the detection of Ancient Silk Textile, particularly relate to a kind of method that dot immuno gold filtration assay measures Ancient Silk Textile.

Background technology

Silk goods are woven by silk and form, it is Chinese nation's crystallization of wisdom, the precious legacy of China, but silk is as a kind of high-molecular organic material, bury in the environment of grave for a long time, will inevitably be subject to the impact of the factors such as oxygen, water, microorganism and degrade, to such an extent as to outward appearance is seriously damaged and rotten, the basic None-identified of naked eyes.Conventional detection method has Infrared spectroscopy, amino acid analysis, liquid-phase chromatographic analysis etc., but because grain rotten silk goods impurity content is many, spectrum elucidation difficulty, length consuming time, sensitivity is low, and can not get rid of the interference of other oroteins, brings certain difficulty to research worker.

Summary of the invention

The technical problem to be solved in the present invention is for this reason: provide a kind of quick, directly perceived, easy Ancient Silk Textile detection method for above-mentioned existing problem.

Adopt following technical scheme: a kind of dot immuno gold filtration assay measures the method for Ancient Silk Textile for this reason, it is characterized in that the step adopted is as follows:

A) getting three specifications is 4 × 3 × 0.6cm plastic caddy, makes a call to the aperture that a diameter is about 0.3-0.8cm respectively in the central authorities of lid; Two-layer adsorptive pads is filled at the lid end; A slice nitrocellulose filter is placed, i.e. NC film above adsorptive pads; On the NC film of then two plastics boxes wherein, point adds the silk fibroin protein solution of 1-2ml 0.5-3mg/l, dries, is labeled as I, II under room temperature; I is positive control, and II is negative control; Get the historical relic sample of 1g silk goods, being dissolved in 100ml PH is 2h in the phosphate-buffered of 7.4, gets the supernatant of 80-100 μ L, is put and is added on NC film, dry, be labeled as III under room temperature;

B), on the NC film below I, II, III aperture, the mass concentration dripping 80-100 μ L is the bovine serum albumen solution of 1%, ensures that solution penetrates into cassette interior;

C), on the NC film below I, III aperture, drip the anti-fibroin albumen antibody-solutions of rabbit that 80-100 μ L 50-100 doubly dilutes, ensure that solution penetrates into cassette interior; II aperture adds the phosphate buffered solution that PH is 7.4, and solution penetrates into cassette interior;

D), on the NC film below I, III aperture, the PH dripping 80-100 μ L is the phosphate-buffered cleansing solution of 7.4, and solution penetrates in box, washs three times altogether;

E), on the NC film below I, II, III aperture, drip the fibroin albumen antibody-solutions of the colloid gold label that 80-100 μ L100-400 doubly dilutes, solution penetrates into cassette interior;

F), on the NC film below I, II, III, the PH dripping 80-100 μ L is the phosphate buffered solution of 7.4, and solution penetrates in box, washing; Wash three times altogether;

G) observations, the NC film of I, III group has red spot occur, the film of II group does not have punctation to occur, illustrates that the sample detected is silk goods.

The present invention adopts miillpore filter as carrier, by antigen or antibody point on film, adds sample to be tested, wash the corresponding antigen of antibody test of rear colloid gold label after closing; Can according to whether carrier occurring redness carries out qualitative analysis.

The invention has the beneficial effects as follows: adopt the method for dot immuno gold filtration assay to detect Ancient Silk Textile, on the one hand, highly sensitive, simple to operate.On the other hand, cost is low, and the reaction time is short.

Embodiment

The step that embodiment 1 adopts is as follows:

A) getting three specifications is 4 × 3 × 0.6cm plastic caddy, makes a call to the aperture that a diameter is about 0.3cm respectively in the central authorities of lid; Two-layer adsorptive pads is filled at the lid end; A slice nitrocellulose filter is placed, i.e. NC film above adsorptive pads; On the NC film of then two plastics boxes wherein, point adds the silk fibroin protein solution of 1ml 0.5mg/l, dries, is labeled as I, II under room temperature; I is positive control, and II is negative control; Get the historical relic sample of 1g silk goods, being dissolved in 100ml PH is 2h in the phosphate-buffered of 7.4, gets the supernatant of 80 μ L, is put and is added on NC film, dry, be labeled as III under room temperature;

B), on the NC film below I, II, III aperture, the mass concentration dripping 80 μ L is the bovine serum albumen solution of 1%, ensures that solution penetrates into cassette interior;

C), on the NC film below I, III aperture, drip the anti-fibroin albumen antibody-solutions of rabbit of 80 μ L 50 times dilution, ensure that solution penetrates into cassette interior; II aperture adds the phosphate buffered solution that PH is 7.4, and solution penetrates into cassette interior;

D) on the NC film below I, III aperture, the PH dripping 80 μ L is the phosphate-buffered cleansing solution of 7.4, and solution penetrates in box, washs three times altogether;

E), on the NC film below I, II, III aperture, drip the fibroin albumen antibody-solutions of the colloid gold label of 80 μ L 100 times dilution, solution penetrates into cassette interior;

F) on the NC film below I, II, III, the PH dripping 80 μ L is the phosphate buffered solution of 7.4, and solution penetrates in box, washing; Wash three times altogether;

G) observations, the NC film of I, III group has red spot occur, the film of II group does not have punctation to occur, illustrates that the sample detected is silk goods.

The step that embodiment 2 adopts is as follows:

A) getting three specifications is 4 × 3 × 0.6cm plastic caddy, makes a call to the aperture that a diameter is about 0.6cm respectively in the central authorities of lid; Two-layer adsorptive pads is filled at the lid end; A slice nitrocellulose filter is placed, i.e. NC film above adsorptive pads; On the NC film of then two plastics boxes wherein, point adds the silk fibroin protein solution of 1.5ml 2mg/l, dries, is labeled as I, II under room temperature; I is positive control, and II is negative control; Get the historical relic sample of 1g silk goods, being dissolved in 100ml PH is 2h in the phosphate-buffered of 7.4, gets the supernatant of 90 μ L, is put and is added on NC film, dry, be labeled as III under room temperature;

B), on the NC film below I, II, III aperture, the mass concentration dripping 90 μ L is the bovine serum albumen solution of 1%, ensures that solution penetrates into cassette interior;

C), on the NC film below I, III aperture, drip the anti-fibroin albumen antibody-solutions of rabbit of 90 μ L 700 times dilution, ensure that solution penetrates into cassette interior; II aperture adds the phosphate buffered solution that PH is 7.4, and solution penetrates into cassette interior;

D) on the NC film below I, III aperture, the PH dripping 90 μ L is the phosphate-buffered cleansing solution of 7.4, and solution penetrates in box, washs three times altogether;

E), on the NC film below I, II, III aperture, drip the fibroin albumen antibody-solutions of the colloid gold label of 90 μ L 300 times dilution, solution penetrates into cassette interior;

F) on the NC film below I, II, III, the PH dripping 90 μ L is the phosphate buffered solution of 7.4, and solution penetrates in box, washing; Wash three times altogether;

G) observations, the NC film of I, III group has red spot occur, the film of II group does not have punctation to occur, illustrates that the sample detected is silk goods.

The step that embodiment 3 adopts is as follows:

A) getting three specifications is 4 × 3 × 0.6cm plastic caddy, makes a call to the aperture that a diameter is about 0.8cm respectively in the central authorities of lid; Two-layer adsorptive pads is filled at the lid end; A slice nitrocellulose filter is placed, i.e. NC film above adsorptive pads; On the NC film of then two plastics boxes wherein, point adds the silk fibroin protein solution of 2ml 3mg/l, dries, is labeled as I, II under room temperature; I is positive control, and II is negative control; Get the historical relic sample of 1g silk goods, being dissolved in 100ml PH is 2h in the phosphate-buffered of 7.4, gets the supernatant of 100 μ L, is put and is added on NC film, dry, be labeled as III under room temperature;

B), on the NC film below I, II, III aperture, the mass concentration dripping 100 μ L is the bovine serum albumen solution of 1%, ensures that solution penetrates into cassette interior;

C), on the NC film below I, III aperture, drip the anti-fibroin albumen antibody-solutions of rabbit of 100 μ L 100 times dilution, ensure that solution penetrates into cassette interior; II aperture adds the phosphate buffered solution that PH is 7.4, and solution penetrates into cassette interior;

D) on the NC film below I, III aperture, the PH dripping 100 μ L is the phosphate-buffered cleansing solution of 7.4, and solution penetrates in box, washs three times altogether;

E), on the NC film below I, II, III aperture, drip the fibroin albumen antibody-solutions of the colloid gold label of 100 μ L 400 times dilution, solution penetrates into cassette interior;

F) on the NC film below I, II, III, the PH dripping 100 μ L is the phosphate buffered solution of 7.4, and solution penetrates in box, washing; Wash three times altogether;

G) observations, the NC film of I, III group has red spot occur, the film of II group does not have punctation to occur, illustrates that the sample detected is silk goods.

Claims (1)

1. dot immuno gold filtration assay measures a method for Ancient Silk Textile, it is characterized in that the step adopted is as follows:

A) getting three specifications is 4 × 3 × 0.6cm plastic caddy, makes a call to the aperture that a diameter is about 0.3-0.8cm respectively in the central authorities of lid; Two-layer adsorptive pads is filled at the lid end; A slice nitrocellulose filter is placed, i.e. NC film above adsorptive pads; On the NC film of then two plastics boxes wherein, point adds the silk fibroin protein solution of 1-2ml 0.5-3mg/l, dries, is labeled as I, II under room temperature; I is positive control, and II is negative control; Get the historical relic sample of 1g silk goods, being dissolved in 100ml PH is 2h in the phosphate-buffered of 7.4, gets the supernatant of 80-100 μ L, is put and is added on NC film, dry, be labeled as III under room temperature;

B), on the NC film below I, II, III aperture, the mass concentration dripping 80-100 μ L is the bovine serum albumen solution of 1%, ensures that solution penetrates into cassette interior;

C), on the NC film below I, III aperture, drip the anti-fibroin albumen antibody-solutions of rabbit that 80-100 μ L 50-100 doubly dilutes, ensure that solution penetrates into cassette interior; II aperture adds the phosphate buffered solution that PH is 7.4, and solution penetrates into cassette interior;

D), on the NC film below I, III aperture, the PH dripping 80-100 μ L is the phosphate-buffered cleansing solution of 7.4, and solution penetrates in box, washs three times altogether;

E), on the NC film below I, II, III aperture, drip the fibroin albumen antibody-solutions of the colloid gold label that 80-100 μ L100-400 doubly dilutes, solution penetrates into cassette interior;

F), on the NC film below I, II, III, the PH dripping 80-100 μ L is the phosphate buffered solution of 7.4, and solution penetrates in box, washing; Wash three times altogether;

G) observations, the NC film of I, III group has red spot occur, the film of II group does not have punctation to occur, illustrates that the sample detected is silk goods.