Human CD117 molecule (CD117) ELISA kit instruction manual - Database & Sql Blog Articles

The domestic authoritative scientific research reagent supplier, specializing in the operation of imported sub-package and original, domestic elisa kit, quality assurance, strict technology, no effect refund return. Welcome to inquire and order. Phone, 60496031 Website: http://

Kit

Can only be used

science

the study

,

Not for medical diagnosis

.

Human CD117 molecule (CD117) ELISA kit

user's Guide

【

Human CD117 molecule (CD117) ELISA kit

Kit name

Human CD117 molecule (CD117) ELISA kit

【

Human CD117 molecule (CD117) ELISA kit

Kit use]

Quantitative detection

people

In serum, plasma and related fluid samples

CD117 molecule (CD117)

The content.

【

Human CD117 molecule (CD117) 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

CD117 molecule (CD117)

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.

CD117 molecule (CD117)

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.

CD117 molecule (CD117)

content.

【

Human CD117 molecule (CD117) ELISA kit

Kit

composition】

1

Enzyme label coated plate

12

hole

×

8

Article

7

Substance night A

6

m

L

2

Standard

:

200ng/ml

0.

6

m

L

8

Substrate night B

6

m

L

3

20

Concentrated concentrated washing solution

2

0

m

L

9

Stop solution

6

m

L

4

Standard dilution

6

m

L

10

Instruction manual

1

Share

5

kind

this

Diluent

6mL

11

Sealing film

1

Zhang

6

Enzyme standard reagent

6

m

L

12

sealed bag

1

One

Remarks:

Standard

Dilute with standard dilutions to: 200, 100, 50, 25, 12.5, 6.25 ng/ml

【

Human CD117 molecule (CD117) 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

【

Human CD117 molecule (CD117) 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 reagent 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

.

【

Human CD117 molecule (CD117) 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.

【

Human CD117 molecule (CD117) 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.

【

Human CD117 molecule (CD117) 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

【

Human CD117 molecule (CD117) ELISA kit

examination range

】

0.1-200ng/ml

【

Human CD117 molecule (CD117) ELISA kit

specification】

96T/box

,

48T/box

【

Human CD117 molecule (CD117) ELISA kit

Storage

2-8 ° C, protected from light and moisture.

【

Human CD117 molecule (CD117) ELISA kit

Validity period]

6 months