CN1118703C - Fast protein adsorbing instrument with applied electric field - Google Patents
Fast protein adsorbing instrument with applied electric field Download PDFInfo
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- CN1118703C CN1118703C CN 99119099 CN99119099A CN1118703C CN 1118703 C CN1118703 C CN 1118703C CN 99119099 CN99119099 CN 99119099 CN 99119099 A CN99119099 A CN 99119099A CN 1118703 C CN1118703 C CN 1118703C
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- liquid groove
- buffering liquid
- silica gel
- casing
- gel sheet
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Abstract
The present invention relates to a fast protein adsorbing instrument with applied electric fields, which is provided with a cover plate and a casing. The present invention is characterized in that the instrument is also provided with a buffer solution slot top board arranged on multiple holes in the casing, a bottom silicon film thereof, a buffer solution slot bottom plate arranged on the multiple holes, a top silicon film thereof, a positive platinum wire electrode and a negative platinum wire electrode. The present invention can effectively absorb multiple parts any ingredient containing protein on a cellulose nitrate film under the action of an electric field, different protein is detected via immune reaction, and the present invention completely overcomes the influence of surfactants. Absorptive speed is as high as only 10 min. The present invention can be directly used for detecting full cell extracts containing surfactants with rapid operation.
Description
Technical field
The present invention relates to a kind of with the instrument of protein adsorption in nitrocellulose filter.
Background technology
The method that detects albumen by immune response is a qualitative leap from precipitation reaction to the enzyme-linked immuno assay (ELISA) that is carrier with 96 hole polystyrene plastic plates, but, because polystyrene plastics plate adsorptive power is limited, the latter's range of application still is subjected to a lot of restrictions, mainly be that antigen purity and surfactant have the greatest impact to testing result, surfactant troublesome more especially.What occurred afterwards is the spot immune analytical approach of carrier with the nitrocellulose filter, because of nitrocellulose filter to the adsorptive power of albumen much larger than the polystyrene plastics plate, can directly test with mixed protein antigen, still, its effect still is subjected to having a strong impact on of surfactant.And ELISA method and spot immune analytical approach are all more time-consuming, and only the antigen adsorption time just reaches 24 hours to 48 hours, and the immunoassay time subsequently also needs more than 2 hours.After this occurred enzyme linked immunological filter analysis (ELIFA) method based on vacuum suction again, this method still is easy to be subjected to the influence of surfactant, and each hole filter velocity is very difficult consistent, and less stable is not widely used so far.
Summary of the invention
The objective of the invention is to overcome the influence of above-mentioned surfactant, a kind of protein adsorption instrument is provided, it can be adsorbed in the albumen that contains any composition on the nitrocellulose filter efficiently, and adsorption rate is fast, detect various albumen by immune response then, accomplish easy and simple to handlely, test is quick.
The objective of the invention is to realize by following technical proposals:
Fast protein adsorbing instrument with applied electric field, has cover plate, casing, its technical scheme is that it also has last buffering liquid groove top board and the bottom silicon film of being located in the casing thereof, last buffering liquid groove base plate and top silica gel sheet thereof, anodal and the platinum electrode negative pole of platinum electrode, nitrocellulose filter, the interior bottom of casing that is used as down buffering liquid groove has the platinum electrode positive pole, the platinum electrode positive pole is made up of a plurality of bending segments, last buffering liquid groove base plate is located at and can be prevented in the casing on the anodal supporter that contacts of itself and platinum electrode, the silica gel sheet fixedly is affixed on the top of buffering liquid groove base plate on this, a plurality of vertical aperture position above the two is corresponding, all according to 96 hole microplate standard ordering of international uniform, in corresponding each aperture in the two position, a cylinder that has central through hole is installed all, each cylinder all passes the silica gel sheet makes its end face be in the outside surface of silica gel sheet, post above-mentioned nitrocellulose filter on the end face of described each cylinder, last buffering liquid groove top board by dismountable web member be fixed in the buffering liquid groove base plate above, the silica gel sheet that is positioned at above-mentioned nitrocellulose filter surface is fixedly posted in last buffering liquid groove top board bottom, a plurality of vertical aperture position above the two is corresponding, all according to 96 hole microplate standard ordering of international uniform, the platinum electrode negative pole is made up of a plurality of bending segments, it is positioned at the zone that described a plurality of vertical aperture forms, and arbitrary vertical aperture is to consistent with the distance of the electrode bending segment of its minimum distance.
In the technique scheme, supporter is a boss of being located at both sides, bottom in the casing.The silica gel sheet of being located at buffering liquid groove top board bottom is spacing fixing by two fixed legs that the diagonal angle on the last buffering liquid groove base plate is provided with respectively with the silica gel sheet of being located at buffering liquid groove base plate top.Above-mentioned dismountable web member is for passing four bolts of four ends of buffering liquid groove top board and last buffering liquid groove base plate angle threaded hole respectively.
The present invention is because it also has the last buffering liquid groove top board be located in the casing and bottom silicon film, last buffering liquid groove base plate and top silica gel sheet, platinum electrode positive pole and platinum electrode negative pole, nitrocellulose filter.It can be adsorbed in the many parts of albumen that contain any composition on the nitrocellulose filter efficiently by effect of electric field like this, detects various albumen by immune response then, has overcome the influence of surfactant fully, and adsorption rate is exceedingly fast, and only needs 10 minutes.Can directly use the full cell extract that contains surfactant and test, swift to operate, only need 45-60 minute from protein adsorption to the immune response overall process.The beneficial effect that instrument of the present invention is had compared with the prior art is as shown in the table:
| Method name | The influence of foreign protein in the antigen | The influence of surfactant in the antigen | Test duration |
| The ELISA method | Must use purifying antigen | Greatly | More than 24 hours |
| The spot immune method | Can use hybrid antigen | Greatly | More than 24 hours |
| The ELIFA method | Can use hybrid antigen | Greatly | Comparatively fast, about 2 hours |
| This instrument | Can use hybrid antigen | There is not influence (result not being produced a bit influence) up to 6.4%SDS | Be exceedingly fast, only need 45-60 minute |
The present invention is further detailed in conjunction with the accompanying drawings below.
Description of drawings
Fig. 1 is a structural representation of the present invention.
Fig. 2 is for being used as down the structural representation of the casing of buffering liquid groove among the present invention.
Fig. 3 is the structural representation of last buffering liquid groove base plate and top silica gel sheet thereof.
Fig. 4 is the structural representation of cylinder.
Fig. 5 is the structural representation of last buffering liquid groove top board and bottom silicon film thereof.
Embodiment
As Fig. 1, Fig. 2, Fig. 3, Fig. 4, shown in Figure 5, the present invention has cover plate 1, casing 4, be located at last buffering liquid groove top board 10 and bottom silicon film 19 thereof in the casing, go up buffering liquid groove base plate 9 and top silica gel sheet 15, platinum electrode positive pole 12 and platinum electrode negative pole 7, nitrocellulose filter.Be used as down that bottoms have platinum electrode positive pole 12 in the casing 4 of buffering liquid groove, platinum electrode positive pole 12 is made up of a plurality of bending segments, this electrode have binding post 2 '.Last buffering liquid groove base plate 9 is located at and can be prevented in the casing that supporter 11 is for being located at the boss of both sides, bottom in the casing 4 on the anodal supporter 11 that contacts of itself and platinum electrode.As shown in Figure 3, Figure 4, silica gel sheet 15 fixedly is affixed on the top of buffering liquid groove base plate 9 on this, a plurality of vertical aperture 14,13 positions above the two are corresponding, all according to 96 hole microplate standard ordering of international uniform, a cylinder 17 that has central through hole all is installed in corresponding each aperture in the two position, each cylinder 17 all passes silica gel sheet 15 makes its end face 16 be in the outside surface of silica gel sheet 15, posts above-mentioned nitrocellulose filter (not drawing among the figure) on the end face 16 of described each cylinder.As Fig. 1, Fig. 3, shown in Figure 5, last buffering liquid groove top board 10 by dismountable web member 5 be fixed in buffering liquid groove base plate 9 above, dismountable web member 5 is held angle threaded hole 5 for passing four of buffering liquid groove top board and last buffering liquid groove base plates ", 5 ' four bolts.The silica gel sheet 19 that is positioned at above-mentioned nitrocellulose filter surface is fixedly posted in last buffering liquid groove top board 10 bottoms, a plurality of vertical aperture 8,18 positions above the two are corresponding, all according to 96 hole microplate standard ordering of international uniform, platinum electrode negative pole 7 is made up of a plurality of bending segments, it is positioned at the zone that described a plurality of vertical aperture 8 forms, and arbitrary vertical aperture 8 is to consistent with the distance of the electrode bending segment of its minimum distance.Platinum electrode negative pole 7 have binding post 3 '.Above-mentionedly be located at the silica gel sheet 19 of buffering liquid groove top board bottom and be located at two fixed legs 6 ' spacing fixing that the silica gel sheet 15 at buffering liquid groove base plate top is provided with by the diagonal angle on the last buffering liquid groove base plate respectively.This moment, two fixed legs 6 ' " and the hole 6 of going up on the buffering liquid groove top board was mated to insert the hole 6 of two silica gel sheets respectively.Binding post 2 ' and 3 ' respectively lead joint pins 2 and 3 below cover plate plug.Above-mentioned cover plate, casing, last buffering liquid groove top board, last buffering liquid groove base plate material are organic glass, and the cylinder material is plastics.
Before the present invention uses, should earlier each parts be taken apart, and then install and use, its method is that the silica gel sheet 15 with last buffering liquid groove base plate top is affixed on its top, makes each vertical aperture 14,13 contrapositions good, inserts cylinder 17 in each aperture.Pour down a certain amount of damping fluid into buffering liquid groove.Carefully will go up buffering liquid groove base plate 9 again and put into down in the buffering liquid groove, liquid make-up makes it flood the aperture at cylinder 17 tops.Carefully will be affixed on cylinder top (being end face), bubble is retained with the nitrocellulose filter that damping fluid soaks.The silica gel sheet 19 of last buffering liquid groove top board bottom is affixed on above-mentioned nitrocellulose filter surface, puts into buffering liquid groove top board 10 again and use bolt 5 fixations.Damping fluid is added in each vertical aperture 8 of going up the buffering liquid groove top board, and additional damping fluid makes it flood the anodal and platinum electrode negative pole of platinum electrode.The protein that will be dissolved in sample buffer add to be gone up in each vertical aperture of buffering liquid groove top board, builds cover plate, energized, and electric current is 200mA effect 10 minutes enough.Take off nitrocellulose filter, carry out immunoassay.
Claims (4)
1, a kind of fast protein adsorbing instrument with applied electric field, has cover plate, casing, it is characterized in that it also has last buffering liquid groove top board (10) and the bottom silicon film of being located in the casing (4) (19) thereof, last buffering liquid groove base plate (9) and top silica gel sheet (15) thereof, platinum electrode positive pole (12) and platinum electrode negative pole (7), nitrocellulose filter, the interior bottom of casing (4) that is used as down buffering liquid groove has platinum electrode positive pole (12), platinum electrode positive pole (12) is made up of a plurality of bending segments, last buffering liquid groove base plate (9) is located at and can be prevented in the casing on the anodal supporter (11) that contacts of itself and platinum electrode, silica gel sheet (15) fixedly is affixed on the top of buffering liquid groove base plate (9) on this, a plurality of vertical aperture (14 above the two, 13) position is corresponding, all according to 96 hole microplate standard ordering of international uniform, a cylinder (17) that has central through hole all is installed in corresponding each aperture in the two position, each cylinder (17) all passes silica gel sheet (15) makes its end face (16) be in the outside surface of silica gel sheet (15), the end face of described each cylinder posts above-mentioned nitrocellulose filter on (16), last buffering liquid groove top board (10) by dismountable web member (5) be fixed in buffering liquid groove base plate (9) above, the silica gel sheet (19) that is positioned at above-mentioned nitrocellulose filter surface is fixedly posted in last buffering liquid groove top board (10) bottom, a plurality of vertical aperture (8 above the two, 18) position is corresponding, all according to 96 hole microplate standard ordering of international uniform, platinum electrode negative pole (7) is made up of a plurality of bending segments, it is positioned at the zone that described a plurality of vertical apertures (8) form, and arbitrary vertical aperture (8) is to consistent with the distance of the electrode bending segment of its minimum distance.
2, absorption instrument according to claim 1 is characterized in that supporter (11) is for being located at the boss of both sides, the interior bottom of casing (4).
3, absorption instrument according to claim 1 is characterized in that the silica gel sheet (15) of being located at the silica gel sheet (19) of buffering liquid groove top board bottom and being located at buffering liquid groove base plate top is spacing fixing by two fixed legs (6 ') that the diagonal angle on the last buffering liquid groove base plate is provided with respectively.
4, absorption instrument according to claim 1 is characterized in that above-mentioned dismountable web member (5) holds four bolts of angle threaded hole (5 ", 5 ') for passing four of buffering liquid groove top board and last buffering liquid groove base plates respectively.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 99119099 CN1118703C (en) | 1999-09-10 | 1999-09-10 | Fast protein adsorbing instrument with applied electric field |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 99119099 CN1118703C (en) | 1999-09-10 | 1999-09-10 | Fast protein adsorbing instrument with applied electric field |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1247983A CN1247983A (en) | 2000-03-22 |
| CN1118703C true CN1118703C (en) | 2003-08-20 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 99119099 Expired - Fee Related CN1118703C (en) | 1999-09-10 | 1999-09-10 | Fast protein adsorbing instrument with applied electric field |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1118703C (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4895676B2 (en) * | 2006-05-17 | 2012-03-14 | ベックマン コールター， インコーポレイテッド | Microplate |
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1999
- 1999-09-10 CN CN 99119099 patent/CN1118703C/en not_active Expired - Fee Related
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| Publication number | Publication date |
|---|---|
| CN1247983A (en) | 2000-03-22 |
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