CN111337660A - Application method of chemiluminescence liquid - Google Patents
Application method of chemiluminescence liquid Download PDFInfo
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- CN111337660A CN111337660A CN201911405206.1A CN201911405206A CN111337660A CN 111337660 A CN111337660 A CN 111337660A CN 201911405206 A CN201911405206 A CN 201911405206A CN 111337660 A CN111337660 A CN 111337660A
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- 238000000034 method Methods 0.000 title claims abstract description 27
- 239000007788 liquid Substances 0.000 title claims abstract description 20
- 239000012528 membrane Substances 0.000 claims abstract description 104
- 239000012224 working solution Substances 0.000 claims abstract description 42
- 238000012546 transfer Methods 0.000 claims abstract description 17
- 238000003384 imaging method Methods 0.000 claims abstract description 16
- 108010001336 Horseradish Peroxidase Proteins 0.000 claims abstract description 15
- 230000009871 nonspecific binding Effects 0.000 claims abstract description 7
- 239000003755 preservative agent Substances 0.000 claims abstract description 4
- 230000002335 preservative effect Effects 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 31
- 238000005406 washing Methods 0.000 claims description 23
- 239000002033 PVDF binder Substances 0.000 claims description 21
- 229920002981 polyvinylidene fluoride Polymers 0.000 claims description 21
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 18
- 239000011534 wash buffer Substances 0.000 claims description 10
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 claims description 8
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 7
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 7
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 7
- 239000006180 TBST buffer Substances 0.000 claims description 5
- 230000000903 blocking effect Effects 0.000 claims description 5
- 238000011534 incubation Methods 0.000 claims description 5
- 238000009736 wetting Methods 0.000 claims description 5
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 claims description 4
- 229920001220 nitrocellulos Polymers 0.000 claims description 4
- 229940056932 lead sulfide Drugs 0.000 claims description 3
- 229910052981 lead sulfide Inorganic materials 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 235000020183 skimmed milk Nutrition 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- SNRUBQQJIBEYMU-UHFFFAOYSA-N Dodecane Natural products CCCCCCCCCCCC SNRUBQQJIBEYMU-UHFFFAOYSA-N 0.000 claims description 2
- 239000007853 buffer solution Substances 0.000 claims description 2
- 238000001962 electrophoresis Methods 0.000 claims description 2
- 229920002401 polyacrylamide Polymers 0.000 claims description 2
- 229910052708 sodium Inorganic materials 0.000 claims description 2
- 239000011734 sodium Substances 0.000 claims description 2
- 238000004520 electroporation Methods 0.000 claims 2
- 239000012530 fluid Substances 0.000 claims 1
- 125000001981 tert-butyldimethylsilyl group Chemical group [H]C([H])([H])[Si]([H])(C([H])([H])[H])[*]C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H] 0.000 claims 1
- 238000007789 sealing Methods 0.000 abstract description 5
- 238000002474 experimental method Methods 0.000 abstract description 3
- 239000000523 sample Substances 0.000 abstract description 3
- 230000010355 oscillation Effects 0.000 abstract 1
- 238000003018 immunoassay Methods 0.000 description 8
- 238000001514 detection method Methods 0.000 description 7
- 230000002285 radioactive effect Effects 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 3
- 239000000020 Nitrocellulose Substances 0.000 description 3
- 238000004020 luminiscence type Methods 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- ILMRJRBKQSSXGY-UHFFFAOYSA-N tert-butyl(dimethyl)silicon Chemical compound C[Si](C)C(C)(C)C ILMRJRBKQSSXGY-UHFFFAOYSA-N 0.000 description 3
- 238000001262 western blot Methods 0.000 description 3
- 238000000636 Northern blotting Methods 0.000 description 2
- 238000002105 Southern blotting Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 229920009405 Polyvinylidenefluoride (PVDF) Film Polymers 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000012631 diagnostic technique Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- XCAUINMIESBTBL-UHFFFAOYSA-N lead(ii) sulfide Chemical compound [Pb]=S XCAUINMIESBTBL-UHFFFAOYSA-N 0.000 description 1
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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- General Health & Medical Sciences (AREA)
- Urology & Nephrology (AREA)
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Abstract
The invention discloses an application method of chemiluminescence liquid, which comprises the following steps: taking off the transferred membrane from the membrane transfer device, and placing the membrane on a shaking table for oscillation sealing for 1 hour at room temperature or 8 hours at the temperature of 2-8 ℃ by utilizing non-specific binding sites on the sealed membrane; fully covering the membrane with ECL working solution, and then incubating for 1 minute at room temperature; absorbing and discarding redundant ECL working solution, placing the film between two layers of preservative films, and removing bubbles between the films; placing the membrane with the protein-adsorbed surface facing upwards in a cassette, taking out the membrane to cover the surface of the membrane in a dark room, and then carrying out exposure operation or scanning imaging on the membrane by using an imaging system. The application method of the chemiluminescent liquid provided by the embodiment can be used for detecting the horseradish peroxidase-labeled probe or antibody in an experiment.
Description
Technical Field
The invention relates to the technical field of biological engineering, in particular to an application method of a chemiluminescent liquid.
Background
At present, the most widely applied detection method in the field of in vitro diagnostic detection reagents for clinical examination is mainly a chemiluminescence immunoassay. The chemiluminescence immunoassay is a higher diagnostic technique developed by clinical immunoassay technology, can carry out quantitative detection, has all the advantages of a radioactive immunoassay and an enzyme-linked immunoassay compared with the radioactive immunoassay and the enzyme-linked immunoassay, and overcomes the defects of the radioactive immunoassay and the enzyme-linked immunoassay. Among them, the chemiluminescent substrate solution used in the chemiluminescent immunoassay method is the most important reagent and is a key substance for limiting chemiluminescent detection. The sensitivity, the luminous value measuring range, the stability and the like of the chemiluminescence substrate solution obviously influence the chemiluminescence immunoassay detection. In the chemiluminescence liquid and the application method thereof in the prior art, the working liquid is used within 10 minutes after being mixed, and the luminescence value and the luminescence gradient are seriously reduced after the working liquid is used for exceeding the time, so that the detection range is narrowed, the linearity is poor, the sensitivity is reduced, and the detection requirement cannot be met.
Disclosure of Invention
The invention aims to provide an application method of chemiluminescent liquid, which comprises the following steps: s1 electrophoresis operation and electric film transfer operation of sodium dodecyl sulfate-polyacrylamide gel, taking off the transferred film from the film transfer device, and placing the film on a shaking table to vibrate and seal for 1 hour at room temperature or 8 hours at the temperature of 2-8 ℃ by using non-specific binding sites on the sealed film; s2, removing the confining liquid, adding primary antibody working solution, and incubating for 1 hour at room temperature by shaking or incubating for 8 hours at 2-8 ℃; s3, washing the membrane for 4-6 times by shaking the membrane washing buffer solution on a shaking table, wherein each time is at least 5 minutes, and washing away the primary antibody working solution; s4 adding a secondary antibody working solution labeled with horseradish peroxidase, and shaking at room temperature to incubate for 1 hour; s5, washing the membrane for 4-6 times by shaking the membrane washing buffer solution on a shaking table, wherein each time lasts for at least 5 minutes, and washing off the secondary antibody working solution; s6 fully covering the membrane with ECL working solution and then incubating for 1 minute at room temperature; s7, absorbing and discarding redundant ECL working solution, placing the film between two layers of preservative films, and removing bubbles between the films; s8, placing the membrane with the protein-adsorbed surface facing upwards in a cassette, taking out the membrane to cover the surface of the membrane in a dark room, and then carrying out exposure operation or scanning imaging on the membrane by using an imaging system.
The exposure operation includes the steps of: s81 is a step of performing a first exposure operation for 1 minute; s82, evaluating the second exposure time according to the result of the first exposure operation; s83 performs a second exposure according to the estimated second exposure time.
The step of placing the side of the membrane which absorbs the protein upwards in a cassette, taking out the membrane to cover the surface of the membrane in a dark room, and then carrying out exposure operation or scanning imaging on the membrane by using an imaging system is completed within 30 minutes after the incubation is completed. The operation of the electrotransformation membrane in S1 is completed by using a transfer membrane, which is a polyvinylidene fluoride membrane or a nitrocellulose membrane. Before the electrotransformation membrane operation in S1, a polyvinylidene fluoride membrane pretreatment step is included, which specifically includes: wetting the polyvinylidene fluoride membrane with methanol; after the polyvinylidene fluoride membrane is changed into a semitransparent state by the wetting of methanol, the polyvinylidene fluoride membrane is wetted by pure water or a transfer buffer solution. And S3, washing the membrane for 4-6 times by shaking the membrane washing buffer solution on a shaking table, wherein each time is at least 5 minutes, and the step of washing off the primary antibody working solution in the primary antibody working solution is carried out, and the adopted washing solution is tert-butyl dimethyl silicon base containing 0.05% of Tween-20 or lead sulfide containing 0.05% of Tween-20. And S1, performing SDS-PAGE operation and electrotransformation operation, taking the transferred membrane off the membrane transferring device, placing the membrane on a shaking table by using non-specific binding sites on the closed membrane, and sealing for 1 hour at room temperature or 8 hours at 2-8 ℃ by shaking, wherein the adopted sealing solution is TBST or PBST containing 5% skimmed milk powder. The primary anti-working solution is a first antibody working solution diluted to 10ng/mL to 1ug/mL by using the confining liquid. The secondary antibody working solution is a secondary antibody working solution which is diluted to 2ng/mL to 100ng/m by the blocking solution and is marked by horseradish peroxidase. The application method of the chemiluminescent solution provided by this embodiment further includes the step of pre-configuring the ECL working solution within 24 hours before the completion of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis operation and the electrotransformation membrane operation.
The application method of the chemiluminescent solution provided in this example can be used to detect horseradish peroxidase (HRP) immobilized on a membrane in an immunoblot, and can detect an antigen of fg to the minimum. The application method of the chemiluminescent solution provided in this embodiment can be used to detect a probe or an antibody labeled with horseradish peroxidase (HRP) in experiments such as Western Blot, northern Blot, Southern Blot, and DotBlot.
Drawings
FIG. 1 is a schematic diagram of the steps of a method for applying a chemiluminescent liquid according to one embodiment of the invention;
fig. 2 is a schematic view of an exposure operation step in the application method of the chemiluminescent liquid according to the first embodiment of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
As shown in fig. 1, the present embodiment provides an application method of a chemiluminescent liquid, including the following steps:
s1 sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) operation and electrotransfer membrane operation, taking the transferred membrane off the membrane transferring device, and placing the membrane on a shaking table to vibrate and seal for 1 hour at room temperature or 8 hours at the temperature of 2-8 ℃ by using non-specific binding sites on the sealed membrane;
s2, removing the confining liquid, adding primary antibody working solution, and incubating for 1 hour at room temperature by shaking or incubating for 8 hours at 2-8 ℃;
s3, washing the membrane for 4-6 times by shaking the membrane washing buffer solution on a shaking table, wherein each time is at least 5 minutes, and washing away the primary antibody working solution;
s4 adding a secondary antibody working solution labeled with horseradish peroxidase (HRP), and incubating for 1 hour at room temperature by shaking;
s5, washing the membrane for 4-6 times by shaking the membrane washing buffer solution on a shaking table, wherein each time lasts for at least 5 minutes, and washing off the secondary antibody working solution;
s6 fully covering the membrane with ECL working solution and then incubating for 1 minute at room temperature;
s7, absorbing and discarding redundant ECL working solution, placing the film between two layers of preservative films, and removing bubbles between the films;
s8, placing the membrane with the protein-adsorbed surface facing upwards in a cassette, taking out the membrane to cover the surface of the membrane in a dark room, and then carrying out exposure operation or scanning imaging on the membrane by using an imaging system.
As shown in fig. 2, the exposure operation includes the steps of:
s81 is a step of performing a first exposure operation for 1 minute;
s82, evaluating the second exposure time according to the result of the first exposure operation;
s83 performs a second exposure according to the estimated second exposure time.
It can be understood by those skilled in the art that the method of applying the chemiluminescent liquid provided by this embodiment can be adjusted to achieve the best exposure effect according to the method of the exposure operation.
The step of placing the side of the membrane which absorbs the protein upwards in a cassette, taking out the membrane to cover the surface of the membrane in a dark room, and then carrying out exposure operation or scanning imaging on the membrane by using an imaging system is completed within 4 hours after the incubation is completed.
Preferably, the step of placing the membrane with the protein-adsorbed side facing upwards in a cassette, taking out the membrane to cover the membrane surface in a dark room, and performing an exposure operation or scanning imaging on the membrane by using an imaging system is completed within 30 minutes after the incubation is completed.
It will be appreciated by those skilled in the art that the membrane incubated by the above steps produces the highest level of luminescence within the first 30 minutes of incubation, which can last up to 4 hours, with a gradual decrease in light signal over time.
The operation of the electrical transfer film in S1 is performed by using a transfer film, where the transfer film is a polyvinylidene fluoride (PVDF) film. It will be appreciated by those skilled in the art that the polyvinylidene fluoride membrane (PVDF) is a solid support commonly used in western blotting. Polyvinylidene fluoride (PVDF) membranes are hydrophobic in nature, and have a large or small pore size, and the binding of the membrane to low molecular weight proteins is more firm as the pore size of the membrane is continuously reduced.
The electrical transfer operation in S1 is performed by using a transfer film, where the transfer film is a nitrocellulose filter membrane (NC). It will be appreciated by those skilled in the art that nitrocellulose membrane (NC) is used as a carrier for the C/T line in colloidal gold test paper and is also where the immune reaction occurs. NC membranes are one of the most important consumables in biological assays.
Before the electrotransformation membrane operation in S1, a polyvinylidene fluoride (PVDF) pretreatment step is included, and the PVDF pretreatment step includes:
wetting a polyvinylidene fluoride membrane (PVDF) with methanol;
after the polyvinylidene fluoride (PVDF) membrane is made translucent by methanol, it is wetted with pure water or a transfer buffer.
And S3 rinsing the membrane for 4-6 times with membrane washing buffer solution on a shaking table, wherein each time is at least 5 minutes, and the step of washing away the primary antibody working solution in the primary antibody working solution is carried out, and the adopted washing solution is tert-butyl dimethyl silicon base (TBS) containing 0.05% of Tween-20.
And S3, washing the membrane for 4-6 times by shaking the membrane washing buffer solution on a shaking table, wherein each time is at least 5 minutes, and the step of washing away the primary antibody working solution in the primary antibody working solution is carried out, and the adopted washing solution is lead sulfide (PBS) containing 0.05% of Tween-20.
And S1, performing SDS-PAGE operation and electrotransformation operation, taking the transferred membrane off the membrane transferring device, placing the membrane on a shaking table by using non-specific binding sites on the closed membrane, and sealing for 1 hour at room temperature or 8 hours at 2-8 ℃ by shaking, wherein the adopted sealing solution is TBST or PBST containing 5% skimmed milk powder.
The TBST is tert-butyl dimethyl silicon base added with TBST; the PBST is lead sulfide added with Tween-20.
The primary anti-working solution is a first antibody working solution diluted to 10ng/mL to 1ug/mL by using the confining liquid.
The secondary antibody working solution is horseradish peroxidase (HRP) labeled secondary antibody working solution diluted to 2ng/mL to 100ng/m by the blocking solution.
The application method of the chemiluminescent liquid provided by this embodiment further comprises the step of pre-configuring the ECL working solution within 24 hours before the SDS-PAGE operation and the electrotransformation membrane operation are completed.
The chemiluminescence solution provided by the embodiment is a Femto-Sensitive ECLSolution, and has a high-sensitivity and non-radioactive chemiluminescence substrate based on Luminol. The application method of the chemiluminescent solution provided in this embodiment can be used to detect a probe or an antibody labeled with horseradish peroxidase (HRP) in experiments such as Western Blot, northern Blot, Southern Blot, and DotBlot.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (10)
1. The application method of the chemiluminescent liquid is characterized by comprising the following steps:
s1 electrophoresis operation and electric film transfer operation of sodium dodecyl sulfate-polyacrylamide gel, taking off the transferred film from the film transfer device, and placing the film on a shaking table to vibrate and seal for 1 hour at room temperature or 8 hours at the temperature of 2-8 ℃ by using non-specific binding sites on the sealed film;
s2, removing the confining liquid, adding primary antibody working solution, and incubating for 1 hour at room temperature by shaking or incubating for 8 hours at 2-8 ℃;
s3, washing the membrane for 4-6 times by shaking the membrane washing buffer solution on a shaking table, wherein each time is at least 5 minutes, and washing away the primary antibody working solution;
s4 adding a secondary antibody working solution labeled with horseradish peroxidase, and shaking at room temperature to incubate for 1 hour;
s5, washing the membrane for 4-6 times by shaking the membrane washing buffer solution on a shaking table, wherein each time lasts for at least 5 minutes, and washing off the secondary antibody working solution;
s6 fully covering the membrane with ECL working solution and then incubating for 1 minute at room temperature;
s7, absorbing and discarding redundant ECL working solution, placing the film between two layers of preservative films, and removing bubbles between the films;
s8, placing the membrane with the protein-adsorbed surface facing upwards in a cassette, taking out the membrane to cover the surface of the membrane in a dark room, and then carrying out exposure operation or scanning imaging on the membrane by using an imaging system.
2. The method of claim 1, wherein the exposing comprises the steps of:
s81 is a step of performing a first exposure operation for 1 minute;
s82, evaluating the second exposure time according to the result of the first exposure operation;
s83 performs a second exposure according to the estimated second exposure time.
3. The method of claim 2, wherein the step of placing the membrane with the protein-adsorbed side facing upwards in a dark box, removing the membrane from the dark box and covering the surface of the membrane with the membrane, and performing the exposure operation or scanning imaging of the membrane using an imaging system is performed within 30 minutes after the incubation is completed.
4. The method of claim 3, wherein the step of electrically transferring the film in S1 is performed by using a transfer film, and the transfer film is a polyvinylidene fluoride film or a nitrocellulose film.
5. The method of claim 4, wherein a polyvinylidene fluoride membrane pretreatment step is included before the electrotransformation membrane operation in S1, specifically including:
wetting the polyvinylidene fluoride membrane with methanol;
after the polyvinylidene fluoride membrane is changed into a semitransparent state by the wetting of methanol, the polyvinylidene fluoride membrane is wetted by pure water or a transfer buffer solution.
6. The method of claim 5, wherein the step of washing away the primary antibody working solution is performed by washing away the primary antibody working solution by shaking the membrane on a shaking table with a membrane washing buffer for 4-6 times, each time for at least 5 minutes, wherein the washing solution is tert-butyldimethylsilyl containing 0.05% Tween-20 or lead sulfide containing 0.05% Tween-20.
7. The method of claim 6, wherein the step of S1 comprises performing SDS-PAGE and electroporation, removing the transferred membrane from the device, blocking the non-specific binding sites on the membrane by shaking for 1 hr at room temperature or 8 hr at 2-8 deg.C, and using 5% skimmed milk powder in TBST or PBST.
8. The method of claim 7, wherein the primary antibody working solution is the primary antibody working solution diluted to 10ng/mL to 1ug/mL with the blocking solution.
9. The method of claim 8, wherein the secondary antibody working solution is horseradish peroxidase-labeled secondary antibody working solution diluted to 2ng/mL to 100ng/m with the blocking solution.
10. The method of claim 8, further comprising the step of pre-configuring the ECL working fluid within 24 hours before the step of performing the sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the step of performing the electroporation.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111929444A (en) * | 2020-08-12 | 2020-11-13 | 四川大学华西医院 | Effective immunoblotting PVDF membrane labeling method, primary anti-incubation method and elution and color development method |
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| US20020132256A1 (en) * | 2000-11-28 | 2002-09-19 | Cavanaugh Philip Gerard | Method for the detection and measurement of hapten-conjugated biological binding entities by western and dot-blot using anti-hapten antibodies |
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| CN110554183A (en) * | 2019-09-17 | 2019-12-10 | 沈阳万类生物科技有限公司 | Hypersensitive ECL chemiluminescent reagent |
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| US20020132256A1 (en) * | 2000-11-28 | 2002-09-19 | Cavanaugh Philip Gerard | Method for the detection and measurement of hapten-conjugated biological binding entities by western and dot-blot using anti-hapten antibodies |
| CN110068568A (en) * | 2018-01-23 | 2019-07-30 | 深圳市泰锐生物科技有限公司 | A kind of chemiluminescence detection kit and its chemical luminescence detection method |
| CN108241065A (en) * | 2018-04-02 | 2018-07-03 | 西安交通大学 | A kind of immune blotting detection method of phosphorylating protein |
| CN108761095A (en) * | 2018-07-27 | 2018-11-06 | 济南大学 | A kind of preparation method and application of the optical electro-chemistry immunosensor based on stannic oxide/stannic disulfide/mesoporous carbonitride |
| CN110554183A (en) * | 2019-09-17 | 2019-12-10 | 沈阳万类生物科技有限公司 | Hypersensitive ECL chemiluminescent reagent |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111929444A (en) * | 2020-08-12 | 2020-11-13 | 四川大学华西医院 | Effective immunoblotting PVDF membrane labeling method, primary anti-incubation method and elution and color development method |
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