Kit
Can only be used
science
the study
,
Not for medical diagnosis
.
Human Adrenergic (EPI) ELISA Kit
user's Guide
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Human Adrenergic (EPI) ELISA Kit
Kit name
Human Adrenergic (EPI) ELISA Kit
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Human Adrenergic (EPI) ELISA Kit
Kit use]
Quantitative detection
people
In serum, plasma and related fluid samples
Adrenaline (EPI)
The content.
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Human Adrenergic (EPI) ELISA Kit
Detection principle
The kit uses a two-antibody two-step sandwich enzyme-linked immunosorbent assay (ELISA). Add standard and sample to be tested to pre-coated
Adrenaline (EPI)
The transparent enzyme label is coated in the plate, and after incubation for a sufficient period of time, the unbound component is washed to remove the enzyme-labeled working solution, and after incubation for a sufficient period of time, the unbound component is washed and removed. Substrate A and B were sequentially added, and the substrate (TMB) was converted to a blue product under the catalysis of horseradish peroxidase (HRP), which turned yellow under the action of acid, and the color depth was in the sample.
Adrenaline (EPI)
The concentration is positively correlated, the OD value is measured at a wavelength of 450 nm, and the sample is calculated according to the OD value of the standard and the sample.
Adrenaline (EPI)
content.
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Human Adrenergic (EPI) ELISA Kit
Kit
composition】
Remarks:
Standard
Dilute with standard dilutions to: 200, 100, 50, 25, 12.5, 6.25 ng/ml
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Human Adrenergic (EPI) ELISA Kit
Reagents and equipment that are needed but not provided
】
1
,
37
°C
temperate box
2,
Standard specification microplate reader
3,
Precision pipettes and disposable tips
4,
Distilled water
5,
Disposable test tube
6,
Absorbent paper
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Human Adrenergic (EPI) ELISA Kit
Steps】
1. Preparation: Remove the kit from the refrigerator and re-balance at room temperature for 30 minutes.
2, dosing: with distilled water will
The 20-fold concentrated washing solution was diluted to the original washing solution.
3. Add standard and sample to be tested: take a sufficient amount
The enzyme label is coated on the plate and fixed on the frame.
Set standard holes separately,
Sample to be tested
this
hole
And blank control holes, record the position of each hole, add standard 50 in the standard hole
μ
L; the sample to be tested is first added to the sample hole to be tested 10
μ
L, plus sample dilution 40
μ
L (ie, the sample is diluted 5 times); blank control wells are not added.
4, incubation: 37
°C water bath or incubator incubation 3
0min.
5, wash the board: discard the liquid, shoot on the absorbent paper
dry
Fill each well with the washing liquid, let stand for 1 min, remove the washing liquid, pat dry on the absorbent paper, and repeat the washing 4 times (you can also use the washing machine to wash the plate according to the instructions).
6. Add the enzyme standard working solution: add the enzyme standard working solution 50 per well.
μ
L, blank control wells are not added.
7. Incubation: Repeat 4 operations.
8. Wash the plate: Repeat the operation of 5.
9, color development: first add color developer A liquid per hole 50
μ
L, then add the developer B liquid 50
μ
L, 37
°C in the dark to develop color for 15min.
10. Termination:
Remove the ELISA plate and terminate each well
Liquid 50μ
L
, stop the reaction
(The color turns from blue to yellow)
.
11, measurement:
Zero the blank hole and use it within 15 minutes after termination.
4
50
Nm wavelength measurement of each hole
Absorbance value
(OD value)
.
12. Calculation:
according to
Calculate the linear regression equation of the standard curve based on the concentration of the standard and the corresponding OD value, and then
kind
this
of
OD
value
,
in
Regression equation
on
Calculate
corresponding
sample
concentration
,
It can also be calculated using various application software.
The final concentration is
actual
Determination
concentration
Multiply by the dilution factor
.
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Human Adrenergic (EPI) ELISA Kit
Sample requirements
1, sample
Cannot contain sodium azide (NaN
3
)
,because
Sodium azide (NaN
3
)Yes
An inhibitor of horseradish peroxidase (HRP).
2,
The specimens should be extracted as soon as possible after collection, and the extraction should be carried out according to the relevant literature. The experiment should be carried out as soon as possible after extraction. If it cannot be tested immediately,
Put the specimen in
-20
°C save, but should
Avoid repeated freezing and thawing
.
3, the sample should be fully centrifuged, no hemolysis and particles.
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Human Adrenergic (EPI) ELISA Kit
Precautions】
1. The experiment is carried out in strict accordance with the operation of the manual. The determination of the experimental results must be based on the reading of the microplate reader.
2,
If the enzyme label is not used up after being opened, it should be immediately placed in a sealed bag and desiccant.
3,
Recommended all
of
Standards, samples
And blank control
Do double detection
, take the average to reduce the experimental error
.
4. Keep in mind that the sample has been diluted 5 times and the calculated result is multiplied by 5 to determine the actual concentration of the sample.
5, the kit quantitative
Surrounded by
0.1-200ng/ml
More than this
The range is calculated from the extension of the standard curve. If it is not used as an accurate quantitative result, please dilute it with a special dilution solution to determine the accurate result (
0.1-200ng/ml
Within the range), multiplying by the total dilution factor is the final concentration of the sample.
6. If the color is too light, the substrate incubation time can be extended appropriately.
7,
for
avoid
Free of cross-contamination,
The standard, sample and blank control should be replaced once for each additional one;
Enzyme label
Common components such as working fluid, sample diluent and substrate, should be cantilevered and must not touch micropores
Not allowed
reuse
Sealing film
.
8. The kit is used during the shelf life. The reagents of different batches must not be mixed.
9. Substrate B is sensitive to light and avoids prolonged exposure to light.
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Human Adrenergic (EPI) ELISA Kit
Summary of operating procedures
Prepare reagents, samples and standards
Add prepared samples and standards,
37
°C reaction 3
0
minute
Wash the plate 4 times, add the enzyme standard reagent,
37
°C reaction 3
0
minute
Wash the plate 4 times, add the coloring solution A, B,
37
°C color development for 15 minutes
Add stop solution
Read within 15 minutes
OD
value
Calculation
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Human Adrenergic (EPI) ELISA Kit
examination range
】
0.1-200ng/ml
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Human Adrenergic (EPI) ELISA Kit
specification】
96T/box
,
48T/box
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Human Adrenergic (EPI) ELISA Kit
Storage
2-8 ° C, protected from light and moisture.
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Human Adrenergic (EPI) ELISA Kit
Validity period]
6 months
Dongguan Yijia Optoelectronics Co., Ltd. , https://www.everbestlcdlcm.com