Mouse Circulating Complex (CIC) ELISA Test Kit

August 15, 2020

Mouse Circulating Complex (CIC) ELISA test kit uses:

The mouse CIC ELISA kit is used for in vitro quantitative detection of CIC in cell supernatant and various body fluids in serum, plasma, tissues, buffers. This kit can detect natural and recombinant CIC. This kit is designed for scientific research, not for clinical diagnosis.

Test principle:

The mouse CIC kit is a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA). Standards with known CIC concentrations and samples with unknown concentrations are added to microwell enzyme plates for detection. Incubate the CIC and biotin-labeled antibodies first. After washing, HRP labeled with avidin was added. After incubation and washing, the unbound enzyme conjugate is removed, and then substrates A and B are added to act simultaneously with the enzyme conjugate. Produce colors. The color depth is proportional to the concentration of CIC in the sample.

Kit contents and preparation

Kit components (stored at 2-8 Â° C)

96-well configuration

48 hole configuration

Preparation

96/48 serving plate

1 board (96T)

Half plate (48T)

Ready-to-use

Plastic membrane cover

1 piece

Half block

Ready-to-use

Standard product: 800umol / L

1 bottle (0.6ml)

1 bottle (0.3ml)

Follow the instructions for sparse

Blank control

1 bottle (1.0ml)

1 bottle (0.5ml)

Ready-to-use

Standard dilution buffer

1 bottle (5ml)

1 bottle (2.5ml)

Ready-to-use

Biotin-labeled anti-CIC antibody

1 bottle (6ml)

1 bottle (3.0ml)

Ready-to-use

Streptavidin-HRP

1 bottle (10ml)

1 bottle (5.0ml)

Ready-to-use

Wash buffer

1 bottle (60ml)

1 bottle (30ml)

Dilute according to the instructions

Substrate A

1 bottle (6.0ml)

1 bottle (3.0ml)

Ready-to-use

Substrate B

1 bottle (6.0ml)

1 bottle (3.0ml)

Ready-to-use

Stop solution

1 bottle (6.0ml)

1 bottle (3.0ml)

Ready-to-use

Mouse Circulating Complex (CIC) ELISA Test Kit

Bring your own materials

1. Distilled water.

2. Sampler: 5ul, 10ul, 50ul, 100ul, 200, 500u, 1000lul.

3. Oscillator and magnetic stirrer etc.

safety

1. Avoid direct contact with the stop solution and substrates A and B. Once exposed to these liquids, rinse with water as soon as possible.

2. Do not eat, drink, smoke or use cosmetics during the experiment.

3. Do not use your mouth to absorb any ingredients in the kit.

Operation notes

1. Reagents should be stored according to the label instructions and returned to room temperature before use. The sparse standards should be discarded and cannot be stored. Mouse Circulating Complex (CIC) ELISA Test Kit

2. The slats not used in the experiment should be immediately returned to the packaging bag, sealed and stored to avoid deterioration.

3. Unused other reagents should be packed or covered. Do not mix reagents of different batches. Use before warranty.

4. Use a disposable pipette tip to avoid cross-contamination, and avoid using a sampler with a metal part when drawing the stop solution and substrates A and B.

5. Use a clean plastic container to prepare the washing solution. Mix all components and samples in the kit thoroughly before use.

6. When washing the enzyme-labeled plate, it should be fully patted dry. Do not put the absorbent paper directly into the enzyme-labeled reaction well to absorb water.

7. Substrate A should evaporate to avoid opening the lid for a long time. Substrate B is sensitive to light and avoid prolonged exposure to light. Avoid contact with hands, it is toxic. The OD value should be read immediately after the experiment is completed.

8. The order of adding reagents should be the same to ensure that all wells are incubated for the same time. Mouse Circulating Complex (CIC) ELISA Test Kit

9. Perform the incubation operation according to the time, the amount and order of the liquid indicated in the instructions.

Sample collection, processing and storage methods

1. Serum-Avoid any cell stimulation during the operation. Use test tubes free of pyrogens and endotoxins. After collecting blood, centrifuge at 1000 Ã— g for 10 minutes to separate the serum and red blood cells quickly and carefully.

2. Plasma ----- EDTA, citrate and heparin plasma can be used for detection. Centrifuge at 1000 Ã— g for 30 minutes to remove particles.

3. The cell supernatant --- 1000 Ã— g centrifuged for 10 minutes to remove particles and polymers.

4. Storage ------ If the sample is not used immediately, it should be divided into small parts and stored at -70 â„ƒ to avoid repeated freezing. If possible, do not use hemolysis or hyperlipidemia. If there are a large number of particles in the serum, centrifuge or filter before testing. Do not thaw at 37 Â° C or higher. Thaw at room temperature and ensure that the sample is thawed evenly and adequately. Mouse Circulating Complex (CIC) ELISA Test Kit

Reagent preparation

1. Standards: Serial dilutions of standards should be prepared during the experiment and cannot be stored. Before dilution, the standard was mixed by shaking. The dilution ratio is as follows:

800 umol / L

(No. 6 standard product)

The original concentration is directly added to 50ul without dilution.

400 umol / L

(No. 5 standard product)

100ul of original standard is added to 100ul of standard dilution

200 umol / L

(Standard 4)

Add 100ul of Standard No. 5 to 100ul of Standard Diluent

100 umol / L

(Standard No. 3)

Add 100ul of Standard No. 4 to 100ul of Standard No. 4

50 umol / L

(Standard No. 2)

Add 100ul of Standard No. 3 to 100ul of Standard Diluent

25 umol / L

(No. 1 standard product)

Add 100ul of Standard No. 2 to 100ul of Standard No. 2

0 umol / L

(Blank control)

The original concentration is directly added to 50ul without dilution.

2. Dilution of washing buffer (20 Ã—): 20-fold dilution with distilled water. Mouse Circulating Complex (CIC) ELISA Test Kit

Steps

1. Before use, mix all reagents thoroughly. Don't make the liquid generate a lot of foam, so as to avoid adding a large number of bubbles during sample addition, which will cause errors in sample addition.

2. The number of slats required is determined by the number of samples to be tested plus the number of standard products. It is recommended to make multiple holes for each standard and blank hole. Each sample is determined according to its own quantity, and those that can use multiple holes can be used for multiple holes.

3. Add 50ul of the diluted standard to the reaction well and 50ul of the sample to be tested in the reaction well. Immediately add 50ul of biotin-labeled antibody. Cover the membrane plate, gently shake and mix, and incubate at 37 Â° C for 1 hour.

4. Shake off the liquid in the wells, fill each well with washing liquid, shake for 30 seconds, shake off the washing liquid, pat dry with absorbent paper. Repeat this operation 3 times. If washing with a plate washer, the number of washes is increased once.

5. Add 80 ul of streptavidin-HRP to each well, gently shake and mix, and incubate at 37 Â° C for 30 minutes.

6. Shake off the liquid in the wells, fill each well with washing liquid, shake for 30 seconds, shake off the washing liquid, pat dry with absorbent paper. Repeat this operation 3 times. If washing with a plate washer, the number of washes is increased once.

7. Add 50ul of substrate A and B to each well, mix gently by shaking, and incubate at 37 Â° C for 10 minutes. Avoid light.

8. Remove the enzyme labeling plate and quickly add 50ul of stop solution. After adding the stop solution, the results should be measured immediately.

9. The OD value of each well was measured at a wavelength of 450 nm. Mouse Circulating Complex (CIC) ELISA Test Kit

Recommended protocol

Standard concentration (umol / L)

800

sample

400

sample

200

sample

100

sample

Result analysis

Taking the absorbance OD value as the ordinate (Y) and the corresponding CIC standard concentration as the abscissa (X), the corresponding curve is made. The CIC content of the sample can be converted from the standard curve to the corresponding concentration according to its OD value.

Limitation

The results of standard No. 6 and above are non-linear, and accurate results cannot be obtained based on this standard curve.

Kit performance

1. Sensitivity: The smallest detection concentration is less than No. 1 standard. Linearity of dilution. The correlation coefficient R between the linear regression of the sample and the expected concentration is 0.990.

2. Specificity: Does not react with other human cytokines.

3. Repeatability: The coefficients of variation within and between plates are less than 10%.

Mouse Circulating Complex (CIC) ELISA Test Kit

Product	Film Faced Plywood, Shuttering Plywood Concrete/Formwork Plywood
Regular Size	1220x2440mm, 915x1830mm, 1250x2500mm, other as requested
Thickness	9mm,12mm,14mm,15mm,16mm, 17mm, 18mm,21mm etc
Film Type	Ordinary Wearable Film, Anti slip film
Film Color	Black, Brown, Red, Green Plastic
Core Material	Poplar, Hardwood, Finger Joint
Glue	MR, E1, E2, WBP/ BWP, melamine, phenolic
Technic	One time hot press, Two Times Hot press
Time of Recycle	2-30times acording to different quality
Logo	Accpet customization
Feature	waterproof, water resistant, Wearable

Product

Film Faced Plywood, Shuttering Plywood
Concrete/Formwork Plywood

Regular Size

1220x2440mm, 915x1830mm, 1250x2500mm, other as requested

Thickness

9mm,12mm,14mm,15mm,16mm, 17mm, 18mm,21mm etc

Film Type

Ordinary Wearable Film, Anti slip film

Film Color

Black, Brown, Red, Green Plastic

Core Material

Poplar, Hardwood, Finger Joint

Glue

MR, E1, E2, WBP/ BWP, melamine, phenolic

Technic

One time hot press, Two Times Hot press

Time of Recycle

2-30times acording to different quality

Logo

Accpet customization

Feature

waterproof, water resistant, Wearable

Film Faced Plywood
Film Faced Plywood,Black Film Faced Plywood,Brown Film Faced Plywood,Rotary Film Faced Plywood,Phenolic Film Faced Plywood
Shouguang Bailing Wood Industry Co., LTD. , http://www.bailingfurniture.com