CN204330771U - A kind of novel speed surveys epidermal stem cells kit - Google Patents
A kind of novel speed surveys epidermal stem cells kit Download PDFInfo
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- CN204330771U CN204330771U CN201520016599.8U CN201520016599U CN204330771U CN 204330771 U CN204330771 U CN 204330771U CN 201520016599 U CN201520016599 U CN 201520016599U CN 204330771 U CN204330771 U CN 204330771U
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- drop bottle
- stem cells
- strips
- epidermal stem
- reactive tank
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Abstract
The utility model discloses a kind of novel speed and survey epidermal stem cells kit.Test-strips, drop bottle is contained with in box body; Described test-strips is perpendicular type test-strips and is provided with multiple reactive tank in machine and transverse direction; Described drop bottle is respectively 10G7 protein antibodies drop bottle, α 6 integrin antibody liquid drop bottle, nitrite ion drop bottle.Once identifying in immunodetection experiment, positive control and negative control can detected while working sample, avoiding the false negative and the false result lost efficacy that may hide due to single part of detection when using immuno-chromatographic test paper strip; The utility model structure is simple, easy to use, and do not need instrument just can detect, detection sensitivity exceeds more than ten times than immuno-chromatographic test paper strip.
Description
Technical field:
The utility model belongs to technical field of immunoassay, and particularly the novel speed of one surveys epidermal stem cells kit.
Background technology:
Skin is the maximum organ of human body, resisting microorganism invasion, ultraviolet radiation and preventing from the loss of moisture, regulate body temperature play an important role, is also one of immune ingredient simultaneously.Except these biological functions, skin maintain people also play a part in person very important.The damage that also can stay in various degree even if the skin injury after birth caused by a variety of causes is healed, still lacks effective treatment means at present.And in fact skin is the tissue that power of regeneration is stronger, the lifelong constantly self of epidermis of skin outer layer, the stem cell continuous proliferation differentiation of its basal part is to replace outer terminally differentiated cells, thus carry out the renewal of institutional framework, the death of okioplast comes off and maintains certain balance with the division of substrate stem cell, and this is the basic demand maintaining normal institutional framework and cell homeostasis.Therefore, Testing and appraisal epidermal stem cells right solution epidermal stem cells how reproduction restraint and regulatory mechanism thereof for promoting that the reparation completely of injured skin function and structure is significant.
Recently, there is experiment in conjunction with α 6 integrin of stem cell surface and another surface marker 10G7 relevant with propagation, epidermal stem cells and of short duration noble cells can be distinguished.Detection finds α 6 positive and the cell of 10G7 feminine gender is in quiescent condition, has very strong proliferation potential, turn out to be epidermal stem cells in cultivating in vitro: and α 6 and the equal positive cell of 10G7 are of short duration noble cellss, and in vitro culture confirms that its multiplication capacity is limited; Its keratin of cell K10 then positive expression of α 6 feminine gender, explanation is terminally differentiated cells after mitosis.Thus α 6 integrin and 10G7 can be utilized to distinguish the epidermal cell of stem cell, of short duration noble cells and differentiation.
Current qualification epidermal stem cells detects respectively multiple marker protein, and do not carry out the positive and negative detection simultaneously, therefore, and qualification
accuracy rate is low,trace routine complexity, length consuming time, be not suitable for quick detection.
The information being disclosed in this background technology part is only intended to increase the understanding to general background of the present utility model, and should not be regarded as admitting or imply in any form that this information structure has been prior art that persons skilled in the art are known.
Utility model content:
The purpose of this utility model is to provide a kind of novel speed to survey epidermal stem cells kit, thus overcomes above-mentioned defect of the prior art.
For achieving the above object, the utility model provides
A kind of novel speed surveys epidermal stem cells kit, comprising: test-strips, drop bottle, box body; Test-strips, drop bottle is contained with in described box body; Described test-strips is perpendicular type test-strips and is provided with multiple reactive tank in machine and transverse direction; Described drop bottle is respectively 10G7 protein antibodies drop bottle, α 6 integrin antibody liquid drop bottle, nitrite ion drop bottle.
Preferably, in technique scheme, perpendicular type test-strips is provided with through damping fluid bag by the negative groove crossed in longitudinal and horizontal intersection, perpendicular type test-strips longitudinally first reactive tank, laterally first reactive tank is through the mixed liquor bag of 10G7 albumen and α 6 integrin by the positive groove crossed, and perpendicular type test-strips longitudinally and transversely remaining reaction groove is sample test groove.
Preferably, in technique scheme, the reactive tank shape in perpendicular type test-strips is circular or square or both combinations.
Preferably, in technique scheme, the square reactive tank length of side is 2-10mm.
Preferably, in technique scheme, the square reactive tank length of side is 5-8mm.
Preferably, in technique scheme, circular reactive tank diameter is 2-10mm.
Preferably, in technique scheme, circular reactive tank diameter is 5-8mm.
Preferably, in technique scheme, 10G7 protein antibodies drop bottle and α 6 integrin antibody liquid drop bottle
In 10G7 protein antibodies and α 6 integrin antibody be all through the antibody of hydrogen oxide enzyme labeling.
Preferably, in technique scheme, nitrite ion drop bottle is built with TMB nitrite ion.
Compared with prior art, the utility model has following beneficial effect:
Once identifying in immunodetection experiment, positive control and negative control can detected while working sample, avoiding the false negative and the false result lost efficacy that may hide due to single part of detection when using immuno-chromatographic test paper strip; The utility model structure is simple, easy to use, and do not need instrument just can detect, detection sensitivity exceeds more than ten times than immuno-chromatographic test paper strip.
Accompanying drawing illustrates:
Fig. 1 is the utility model structural representation;
Main Reference Numerals illustrates:
1-perpendicular type test-strips, 2-10G7 protein antibodies drop bottle, 3-α 6 integrin antibody liquid drop bottle, 4-nitrite ion drop bottle, 5-box body.
Embodiment:
Below embodiment of the present utility model is described in detail, but is to be understood that protection domain of the present utility model not by the restriction of embodiment.
Clearly represent unless otherwise other, otherwise in whole instructions and claims, term " comprise " or its conversion as " comprising " or " including " etc. by be understood to include the element of stating or ingredient, and do not get rid of other element or other ingredient.
A kind of novel speed surveys epidermal stem cells kit, comprising: test-strips, drop bottle, box body; Test-strips, drop bottle is contained with in described box body; Described test-strips is perpendicular type test-strips 1 and is provided with multiple reactive tank in machine and transverse direction; Described drop bottle is respectively 10G7 protein antibodies drop bottle 2, α 6 integrin antibody liquid drop bottle 3, nitrite ion drop bottle 4.
Embodiment 1
A, prepare kit
As shown in Figure 1, polystyrene plastics is selected to be injection molded as perpendicular type test-strips 1.The square reactive tank (being labeled as Y) of the intersection in length and breadth of described perpendicular type test-strips 1 is as negative groove, and its length of side is 8mm, and its adjacent vertical and horizontal respectively have 3 circular reactive tanks, and circular reactive tank diameter is 8mm.PBS damping fluid 100ul is added in negative groove, longitudinally first reactive tank (be labeled as A) adjacent with this negative groove and first horizontal reactive tank (being labeled as G) are as positive groove, in positive groove, respectively add 100ul 10G7 albumen and α 6 integrate white mixed liquor (10G7 albumen, the each 2ug/ml of α 6 integrin), room temperature leaves standstill washing trough after 4 hours, then at negative groove, respectively add 200ul in positive groove to contain the PBS damping fluid of 2%BSA (all the other each reactive tanks are left intact, fix when giving over to detection cell to be measured with) room temperature leave standstill close 2 hours, then dry, aluminium foil bag vacuum seal is used after 40 DEG C of oven dry, be placed in refrigerator for subsequent use.
By the epidermal stem cells mark that hydrogen peroxidase (HRP) marks---10G7 protein antibodies and α 6 integrin antibody are released to 1ug/ml with 1% bovine serum albumin-PBS damping fluid respectively, after aseptic filtration, be contained in 10G7 protein antibodies drop bottle 2,3 li, the α 6 integrin antibody liquid drop bottle of 2ml-capacity again by every bottle of 0.5ml respectively, be placed in refrigerator for subsequent use.
TMB is dissolved in 50mM citrate buffer solution (pH4.5), degree of thickening is to 0.2mg/ml, add hydrogen peroxide to 0.03% (this TMB-hydrogen peroxide solution is hydrogen peroxidase nitrite ion) again, be divided in the drop bottle of 3ml-capacity by every bottle of 2ml again, be contained in 4 li, nitrite ion drop bottle after aseptic filtration, be placed in refrigerator for subsequent use.
B, detection
1) cell liquid to be checked prepares: the epidermal stem cells and the fibrocyte PBS that get in vitro culture respectively adjust cell density to 50 ten thousand/milliliter, for subsequent use;
2) cell to be measured is fixed: get the perpendicular type test-strips described in step A 1 one, be respectively each reactive tank marking: negative groove is Y, its longitudinal first is A (positive groove), longitudinally the 2nd, 3 is respectively A2, A3, laterally first is G (positive groove), and the 2nd, 3 is respectively G2, G3; Step 1 is added respectively in A2 groove and G2 groove) the middle epidermal stem cells liquid 100ul allocated, in A3 and G3 groove, add step 1) the middle fiber finer cytosol 100ul allocated; Then in A2, G2, A3, G3 groove, add 40% acetaldehyde of 100ul respectively again, room temperature left standstill after 2 hours, and gentle aspiration supernatant discards, (now cell has been fixed on reactive tank); Then in A2, G2, A3, G3 groove, respectively add the PBS damping fluid that 200ul contains 2%BSA, room temperature leaves standstill closes 2 hours, then dries, for subsequent use;
3) the 10G7 protein antibodies liquid that each dropping 3 (about 100ul) hydrogen peroxidases (HRP) mark in reactive tank Y, G, G2, G3 respectively, each α 6 integrin antibody liquid dripping 3 (about 100ul) hydrogen peroxidases (HRP) and mark in reactive tank Y, A, A2, A3; Left at room temperature 60 minutes after light rolling;
4) wash: outwell liquid in reactive tank and rinse reactive tank 5 times by filtered water, after each flushing, reactive tank is fallen dry again with the globule in thieving paper removing reactive tank;
5) develop the color: in each reactive tank, add 3 (about 100ul) nitrite ions, light shake test-strips color change in observing response groove, until there is blue color clearly to occur in negative reaction groove;
6) interpretation of result: the shade degree of more each reactive tank (positive control, negative control and each sample) is to determine that this test sample is positive or negative.
This test findings shows: negative groove (Y) is colour developing not, positive groove (A and G) is all developed the color, add the A2 groove colour developing (illustrating containing epidermal stem cells mark α 6 integrin) of epidermal stem cells and G2 groove does not develop the color (illustrating not containing 10G7 albumen), add the G3 groove colour developing (illustrating containing the special 10G7 albumen of non-epidermal stem cells) without colour developing (illustrating not containing epidermal stem cells mark α 6 integrin) of fibrocellular A3 groove.This experiment results proved epidermal stem cells quick testing reagent of the present invention box may be used for Testing and appraisal epidermal stem cells.
The aforementioned description to concrete exemplary of the present utility model is to illustrate and the object of illustration.These descriptions not want the utility model to be defined as disclosed precise forms, and obviously, according to above-mentioned instruction, can much change and change.The object selected exemplary embodiment and describe is to explain certain principles of the present utility model and practical application thereof, thus those skilled in the art can be realized and utilize various different exemplary of the present utility model and various different selection and change.Scope of the present utility model is intended to limited by claims and equivalents thereof.
Claims (9)
1. novel speed surveys an epidermal stem cells kit, it is characterized in that: comprise, test-strips, drop bottle, box body; Test-strips, drop bottle is contained with in described box body; Described test-strips is perpendicular type test-strips and is provided with multiple reactive tank in machine and transverse direction; Described drop bottle is respectively 10G7 protein antibodies drop bottle, α 6 integrin antibody liquid drop bottle, nitrite ion drop bottle.
2. novel speed according to claim 1 surveys epidermal stem cells kit, it is characterized in that: described perpendicular type test-strips is provided with through damping fluid bag by the negative groove crossed in longitudinal and horizontal intersection, perpendicular type test-strips longitudinally first reactive tank, laterally first reactive tank is through the mixed liquor bag of 10G7 albumen and α 6 integrin by the positive groove crossed, and perpendicular type test-strips longitudinally and transversely remaining reaction groove is sample test groove.
3. novel speed according to claim 1 surveys epidermal stem cells kit, it is characterized in that: the reactive tank shape in described perpendicular type test-strips is circular or square or both combinations.
4. novel speed according to claim 3 surveys epidermal stem cells kit, it is characterized in that: the described square reactive tank length of side is 2-10mm.
5. novel speed according to claim 4 surveys epidermal stem cells kit, it is characterized in that: the described square reactive tank length of side is 5-8mm.
6. novel speed according to claim 3 surveys epidermal stem cells kit, it is characterized in that: described circular reactive tank diameter is 2-10mm.
7. novel speed according to claim 6 surveys epidermal stem cells kit, it is characterized in that: described circular reactive tank diameter is 5-8mm.
8. novel speed according to claim 1 surveys epidermal stem cells kit, it is characterized in that: the 10G7 protein antibodies in described 10G7 protein antibodies drop bottle and α 6 integrin antibody liquid drop bottle and α 6 integrin antibody are all through the antibody of hydrogen oxide enzyme labeling.
9. novel speed according to claim 1 surveys epidermal stem cells kit, it is characterized in that: described nitrite ion drop bottle is built with TMB nitrite ion.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201520016599.8U CN204330771U (en) | 2015-01-07 | 2015-01-07 | A kind of novel speed surveys epidermal stem cells kit |
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| CN201520016599.8U CN204330771U (en) | 2015-01-07 | 2015-01-07 | A kind of novel speed surveys epidermal stem cells kit |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107907397A (en) * | 2017-11-23 | 2018-04-13 | 苏州泽科生物科技有限公司 | Collagenous fibres horse pine trichrome stain kit and preparation method thereof and colouring method |
| CN109187943A (en) * | 2018-08-24 | 2019-01-11 | 四川新健康成生物股份有限公司 | The preparation method of anti-interference coating in a kind of anti-interference reagent cup and reagent cup |
-
2015
- 2015-01-07 CN CN201520016599.8U patent/CN204330771U/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107907397A (en) * | 2017-11-23 | 2018-04-13 | 苏州泽科生物科技有限公司 | Collagenous fibres horse pine trichrome stain kit and preparation method thereof and colouring method |
| CN109187943A (en) * | 2018-08-24 | 2019-01-11 | 四川新健康成生物股份有限公司 | The preparation method of anti-interference coating in a kind of anti-interference reagent cup and reagent cup |
| CN109187943B (en) * | 2018-08-24 | 2021-07-30 | 四川新健康成生物股份有限公司 | Anti-interference reagent cup and preparation method of anti-interference coating in reagent cup |
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| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150513 Termination date: 20210107 |