CN209226886U - A kind of auxiliary device of phosphorylated polypeptide enrichment - Google Patents
A kind of auxiliary device of phosphorylated polypeptide enrichment Download PDFInfo
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- CN209226886U CN209226886U CN201821917095.3U CN201821917095U CN209226886U CN 209226886 U CN209226886 U CN 209226886U CN 201821917095 U CN201821917095 U CN 201821917095U CN 209226886 U CN209226886 U CN 209226886U
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- auxiliary device
- enrichment
- phosphorylated polypeptide
- suction nozzle
- syringe
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Abstract
A kind of auxiliary device of phosphorylated polypeptide enrichment, including circular platform type polypropylene pipette tips, use for laboratory mineral wool, enrichment microballon, homalocephalus clip, syringe, use for laboratory mineral wool is kneaded into a branch of, fill in the thinner one end of polypropylene pipette tips, using the thinner one end of homalocephalus clamp suction nozzle to fix mineral wool, enrichment microballon is filled in suction nozzle above mineral wool.Syringe adapter is circular platform type, and central angle is identical as the central angle size of polypropylene pipette tips, and syringe adapter outer wall is coated with one layer of silica gel.Polypropylene pipette tips volume is 200 microlitres, long 83mm, and thinner one end hole sucting diameter is 0.2mm.Syringe is 50 milliliters of syringes.The auxiliary device is convenient for the enrichment of micro phosphorylated polypeptide, while can reduce the loss of phosphorylated polypeptide.
Description
Technical field
The utility model relates to the enrichment experiment field of phosphorylated polypeptide, especially a kind of auxiliary of phosphorylated polypeptide enrichment
Device.
Background technique
Protein phosphorylation posttranslational modification has great importance in life entity, and the phosphorylation of protein can change
The property of protein catalytic group, the conformation for changing protein change the positioning of protein in the cell etc., it be it is intracellular
The most quick universal signal transduction mode known.The phosphorylation and dephosphorylation of protein almost regulate and control all in life entity
Life process, increment, development, differentiation and apoptosis, the metabolism of life entity, nervous activity, muscular movement including cell etc.
Deng.The research of phosphorylating protein needs to be enriched with the peptide molecule of phosphorylation, conventional enrichment experiment be will be enriched with it is micro-
Pearl and sample solution, cleaning solution, substitutional solution, elution solution etc. carry out mixing concussion in EP pipe and are incubated for, utilize after centrifugation
Liquid-transfering gun manually separates supernatant fluid and microballon, and since bead particles are small, light weight, manually separation is frequently not band
The loss that part microballon causes sample has been walked, has been exactly remaining too many cleaning solution, leads to the residual of non-phosphorylated polypeptide, make to be enriched with
Efficiency decline.Auxiliary device provided by the utility model is that bead particles and a variety of are separated using a kind of novel filter device
Solution.
Utility model content
In order to solve the problems in the existing technology, the utility model provides a kind of auxiliary dress of phosphorylated polypeptide enrichment
It sets, can be improved the separative efficiency of enrichment microballon and cleaning solution, while the loss of enrichment microballon can be reduced, to reduce phosphoric acid
Change loss of the polypeptide in enrichment process.
A kind of auxiliary device of phosphorylated polypeptide enrichment, including circular platform type polypropylene pipette tips, use for laboratory mineral wool, enrichment
Microballon, homalocephalus clip, syringe, use for laboratory mineral wool are kneaded into a branch of, fill in the thinner one end of polypropylene pipette tips, use is flat
To fix mineral wool, the enrichment microballon is filled in suction nozzle above mineral wool for head thinner one end of clamp suction nozzle.
There is lines in the collet inside of a kind of auxiliary device of above-mentioned phosphorylated polypeptide enrichment, the homalocephalus clip, described
Lines is perpendicular to suction nozzle.
A kind of auxiliary device of above-mentioned phosphorylated polypeptide enrichment, the thicker one end outer wall of the suction nozzle uniformly stretch out at least
Two fixed links.
A kind of auxiliary device of above-mentioned phosphorylated polypeptide enrichment, it is solid that the thicker one end outer wall of the suction nozzle extends a circle
Determine edge.
A kind of auxiliary device of above-mentioned phosphorylated polypeptide enrichment, the syringe adapter are circular platform type, central angle with
The central angle size of polypropylene pipette tips is identical, and syringe adapter outer wall is coated with one layer of silica gel.
A kind of auxiliary device of above-mentioned phosphorylated polypeptide enrichment, the thicker one end of the polypropylene pipette tips is hard, thinner
One end is soft.
A kind of auxiliary device of above-mentioned phosphorylated polypeptide enrichment, the polypropylene pipette tips volume is 200 microlitres, long
83mm, thinner one end hole sucting diameter are 0.2mm.
A kind of auxiliary device of above-mentioned phosphorylated polypeptide enrichment, the syringe are 50 milliliters of syringes.
The advantages of the utility model:
(1) this auxiliary device can efficiently be totally separated from microballon and cleaning solution, can almost retain microballon and not damage
It loses and is nearly completely separated liquid and do not remain.
(2) 200 microlitres of sample can be at most accommodated using the suction nozzle.
(3) special suction nozzle is carefully grown, and a small amount of enrichment microballon can be made to be filled in longer one section of region, so that cleaning
It can more fully be contacted with microballon with elution solution, so that cleaning and elution effect are improved.
(4) there is the other mineral wool of laboratory level very very thin diameter can be enriched with microballon with high efficiency filter.
(5) mineral wool of speciality has few adsorptivity to guarantee that sample does not lose.
(6) time that liquid flows through can be controlled in time using syringe pressure, to guarantee that phosphated peptide section obtains sufficiently
Affine or elution time.
(7) present apparatus is suitable for IMAC microballon, TiO2The separation of solid and liquid of the various enriching and purifying microballons such as microballon.
Detailed description of the invention
Fig. 1 is the sucker structure schematic diagram of the utility model embodiment 1；
Fig. 2 is the sucker structure schematic diagram of the utility model embodiment 2；
Fig. 3 is the sucker structure schematic diagram of the utility model embodiment 3；
Fig. 4 is the structural schematic diagram that suction nozzle fills emitter.
1, suction nozzle in figure, 101, enrichment microballon, 102, mineral wool, 103, fixed link, 104, fixed edge, 2, syringe,
201, connector, 202, scale.
Specific embodiment
[embodiment 1]
A kind of auxiliary device of phosphorylated polypeptide enrichment, including circular platform type polypropylene pipette tips 1, use for laboratory mineral wool 102,
It is enriched with microballon 101, homalocephalus clip, syringe 2, use for laboratory mineral wool 102 is kneaded into a branch of, it is relatively thin to fill in polypropylene pipette tips 1
One end, using the thinner one end of homalocephalus clamp suction nozzle to fix mineral wool, the enrichment microballon is filled in glass in suction nozzle
Above glass cotton.The present apparatus is suitable for IMAC microballon, TiO2The separation of solid and liquid of the various enriching and purifying microballons such as microballon.The mineral wool
English name be Glass Wool, article No. and specification are 20411, are produced by Sigma-Aldrich company.
[embodiment 2]
Based on embodiment 1, the thicker one end outer wall of the suction nozzle 1 uniformly stretches out at least two fixed links 103, can be two
Root, three, four etc..Fixed link is conducive to for suction nozzle to be fixed on shelf etc., facilitates placement.
[embodiment 3]
Based on embodiment 1, it is fixed along 104 that the thicker one end outer wall of the suction nozzle 1 extends a circle.Compared to embodiment 2
Fixed link 103, it is fixed along have the advantages that contact area more greatly, be physically easier to perform and transport when will not break packaging bag.
[embodiment 4]
Based on any of the above embodiment, the auxiliary device of the phosphorylated polypeptide enrichment, the syringe adapter 201 is circle
Bench-type, central angle is identical as the central angle size of polypropylene pipette tips, and 201 outer wall of syringe adapter is coated with one layer of silica gel.Note
Scale 202 is additionally provided on emitter connector.Similarly, scale can also be set on polypropylene pipette tips.Due in use, syringe needs to add
Pressure makes solution slowly flow across enrichment microballon, so the central angle of syringe adapter is identical as the central angle size of suction nozzle, makes to inject
Device can be bonded with suction nozzle, be conducive to sealing consequently facilitating pressurization；The syringe adapter outer wall is coated with one layer of silica gel, can make to inject
Device connector is more bonded with suction nozzle, further increases sealing effect.
[embodiment 5]
Based on any of the above embodiment, the following are preferred features, and one can be selected with any of the above embodiment or is selected more
Combination.
Preferably, there is lines on the inside of the collet of the homalocephalus clip, the lines is perpendicular to suction nozzle.Lines is perpendicular to suction nozzle
Microballon can be prevented to be carried over.
Preferably, the thicker one end of the polypropylene pipette tips is hard, and thinner one end is soft.Thicker one end is hard, convenient for hand-held
With fixation, thinner one end is soft, is conducive to clamp.
Preferably, the polypropylene pipette tips volume is 200 microlitres, long 83mm, and thinner one end hole sucting diameter is 0.2mm.
Preferably, the syringe is 50 milliliters of syringes.
Enrichment method is as follows:
1. taking out 20 μ L IMAC slurry to low adsorption EP pipe sucks supernatant after centrifugation precipitates beads.
2. dissolving the peptide fragment sample drained with 50 μ L 60%ACN/1%AA, supernatant solution is moved into equipped with beads's
In EP pipe, blown several times with rifle suction.
3. EP pipe is placed on metal oscillator, room temperature acutely shakes 1500rpm, 1h.
4. the solution containing Beads is added to together in special phosphorylation enrichment auxiliary device.
5. flowing out solution with the flow velocity of 20 μ L/min using the pressurization of 50mL syringe.
6. 50 μ L 0.1M NaCl/25%ACN/0.1%acetic acid are added, keep solution slow using syringe pressurization
Slug flow crosses enrichment microballon, and repeated washing is primary.
7. 50 μ L 60%ACN/1%AA are added, solution is set to slowly flow across enrichment microballon using syringe pressurization, and repeat
Cleaning is primary.
8. 50 μ LH2O are added, solution is set to slowly flow across enrichment microballon using syringe pressurization, and repeated washing is primary.
9. 40 μ L 6%NH are added3H2O makes solution flow through enrichment microballon with the flow velocity of 20 μ L/min using syringe pressurization,
Collect the liquid of outflow.
10. repeating step 9 twice, the phosphorylated polypeptide solution of 120 μ L is collected altogether.
11. the solution concentrate drying of acquisition can be carried out upper machine testing with mass spectrum.
Claims (8)
1. a kind of auxiliary device of phosphorylated polypeptide enrichment, which is characterized in that including circular platform type polypropylene pipette tips, use for laboratory glass
Glass cotton, enrichment microballon, homalocephalus clip, syringe, use for laboratory mineral wool are kneaded into a branch of, fill in polypropylene pipette tips thinner one
End, using the thinner one end of homalocephalus clamp suction nozzle to fix mineral wool, the enrichment microballon is filled in mineral wool in suction nozzle
Top.
2. a kind of auxiliary device of phosphorylated polypeptide enrichment according to claim 1, which is characterized in that the homalocephalus clip
Collet on the inside of have lines, the lines is perpendicular to suction nozzle.
3. a kind of auxiliary device of phosphorylated polypeptide enrichment according to claim 1, which is characterized in that the suction nozzle is thicker
One end outer wall uniformly stretch out at least two fixed links.
4. a kind of auxiliary device of phosphorylated polypeptide enrichment according to claim 1, which is characterized in that the suction nozzle is thicker
One end outer wall extend the fixed edge of a circle.
5. a kind of auxiliary device of phosphorylated polypeptide enrichment according to claim 1, which is characterized in that the syringe connects
Head is circular platform type, and central angle is identical as the central angle size of polypropylene pipette tips, and syringe adapter outer wall is coated with one layer of silica gel.
6. a kind of auxiliary device of phosphorylated polypeptide enrichment according to claim 1, which is characterized in that the polypropylene is inhaled
The thicker one end of head is hard, and thinner one end is soft.
7. a kind of auxiliary device of phosphorylated polypeptide enrichment according to claim 1, which is characterized in that the polypropylene is inhaled
Head volume is 200 microlitres, long 83mm, and thinner one end hole sucting diameter is 0.2mm.
8. a kind of auxiliary device of phosphorylated polypeptide enrichment according to claim 1, which is characterized in that the syringe is
50 milliliters of syringes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201821917095.3U CN209226886U (en) | 2018-11-21 | 2018-11-21 | A kind of auxiliary device of phosphorylated polypeptide enrichment |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201821917095.3U CN209226886U (en) | 2018-11-21 | 2018-11-21 | A kind of auxiliary device of phosphorylated polypeptide enrichment |
Publications (1)
| Publication Number | Publication Date |
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| CN209226886U true CN209226886U (en) | 2019-08-09 |
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| CN201821917095.3U Active CN209226886U (en) | 2018-11-21 | 2018-11-21 | A kind of auxiliary device of phosphorylated polypeptide enrichment |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110922448A (en) * | 2019-12-11 | 2020-03-27 | 谱度众合(武汉)生命科技有限公司 | Phosphorylated peptide enrichment device and method |
-
2018
- 2018-11-21 CN CN201821917095.3U patent/CN209226886U/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110922448A (en) * | 2019-12-11 | 2020-03-27 | 谱度众合(武汉)生命科技有限公司 | Phosphorylated peptide enrichment device and method |
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