Cattle Growth Hormone (GH) ELISA Kit
This reagent is for research use only Specimens: serum or plasma
Test principle:
The GH kit is a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA). Standards with known GH concentrations and samples with unknown concentrations are added to microwell enzyme plates for detection. Incubate GH 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 GH in the sample.
Kit content and preparation:
Kit ingredients
96-well configuration
48 hole configuration
96/48 serving plate
1 board (96T)
Half block (48T)
Plastic membrane cover
1 piece
Half block
Standard product: 40ng / ml
1 bottle (1.0ml)
1 bottle (0.5ml)
Blank control
1 bottle (1.0ml)
1 bottle (0.5ml)
Standard dilution buffer
1 bottle (8.0ml)
1 bottle (4.0ml)
Biotin-labeled anti-GH antibody
1 bottle (8.0ml)
1 bottle (4.0ml)
Streptavidin-HRP
1 bottle (12ml)
1 bottle (6.0ml)
Wash buffer
1 bottle (20ml)
1 bottle (10ml)
Substrate A
1 bottle (6.0ml)
1 bottle (3.0ml)
Substrate B
1 bottle (6.0ml)
1 bottle (3.0ml)
Stop solution
1 bottle (6.0ml)
1 bottle (3.0ml)
Bring your own materials:
1. Distilled water.
2. Sampler: 5ul, 10ul, 50ul, 100ul, 200, 500ul, 1000ul.
3. Oscillator and magnetic stirrer etc.
4.
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 × 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 × g for 30 minutes to remove particles.
3. Cell supernatant --- 1000 × g centrifuged for 10 minutes to remove particles and polymer.
4. Tissue homogenate ----- Mash the tissue with appropriate amount of saline. Centrifuge at 1000 × g for 10 minutes, take the supernatant
5. Storage ------ If the sample is not used immediately, it should be divided into small parts and stored at -70 ℃ 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 ° C or higher. Thaw at room temperature and ensure that the sample is thawed evenly and adequately.
Operation notes:
l 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.
l The slats not used in the experiment should be immediately returned to the packaging bag, sealed and stored to avoid deterioration.
l Any unused reagents should be packed or covered. Do not mix reagents of different batches. Use before warranty.
l Use disposable pipette tips to avoid cross-contamination. Avoid using a sampler with a metal part when drawing the stop solution and the substrates A and B.
l Use a clean plastic container to prepare the washing solution. Mix all components and samples in the kit thoroughly before use.
l 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.
l The order of adding reagents should be the same to ensure the same incubation time of all reaction wells.
l Carry out the incubation operation according to the time, the amount and order of the liquid indicated in the manual.
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.
Preparation of reagents:
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:
40
ng / ml
(No. 6 standard product)
The original concentration is directly added to 50ul without dilution.
20
ng / ml
(No. 5 standard product)
100ul of original standard is added to 100ul of standard dilution
10
ng / ml
(Standard 4)
Add 100ul of Standard No. 5 to 100ul of Standard Diluent
5.0
ng / ml
(Standard No. 3)
Add 100ul of Standard No. 4 to 100ul of Standard No. 4
2.5
ng / ml
(Standard No. 2)
Add 100ul of Standard No. 3 to 100ul of Standard Diluent
1.25 ng / ml
(No. 1 standard product)
Add 100ul of Standard No. 2 to 100ul of Standard No. 2
0 ng / ml
(Blank control)
The original concentration is directly added to 50ul without dilution.
2. Dilution of washing buffer (50 ×): 50-fold dilution with distilled water.
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 cytokines.
3. Repeatability: The coefficients of variation within and between plates are less than 10%.
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, mix gently by shaking, and incubate at 37 ° C for 45 minutes.
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 4 times. If washing with a plate washer, the number of washes is increased once.
5. Add 100ul of streptavidin-HRP to each well, gently shake to mix, and incubate at 37 ° 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 4 times. If washing with a plate washer, the number of washes is increased once.
7. Add 50ul each of substrate A and B to each well, gently shake to mix, and incubate at 37 ° C for 5 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.
Judgment and analysis of results:
1. Instrument value: read the OD value of each well on a microplate reader with a wavelength of 450nm
2. Take the absorbance OD value as the ordinate (Y) and the corresponding GH standard concentration as the abscissa (X), and make the corresponding curve. The GH content of the sample can be converted from the standard curve to the corresponding concentration according to its OD value. Multiply by the dilution factor; or use the standard concentration and OD value to calculate the regression equation of the standard curve, substitute the OD value of the sample into the equation, calculate the sample concentration, and multiply by the dilution factor to obtain the actual concentration of the sample.
3. Range of detection value: 0-40ng / ml
4. Sensitivity: 0.1 ng / ml
Lip glosses are intended to add shine to the Lips, and may also add a tint of color, as well as being scented or flavored.
Lip Gloss was invented by Max Factory in 1930. He wanted to create a lip product that would make lips shiny and glossy for films.Factor created makeup for the movie industry. He developed makeup specifically for actresses starring in black and white films. Women were inspired by movie actresses and they also wanted makeup. The first commercially available lip gloss was Max factory's X-Rated, launched in 1932.The original formula was sold up until 2003, when procter and gamble retired the product.
Lip gloss is usually used as a cosmetic, however some offer moisturizing benefits or protection from the elements and other natural causes. Lip gloss containing sunscreen was first advertised by actress lillian gish
In 1973, Bonne Bell introduced the first flavored lip gloss, Lip smackers. Lip Smackers were, and still are, popular among young teenagers. Initially Lip Smackers came in two sizes: small and big. The small ones could be kept in the pocket and the big ones had a rope to hang around the neck. It was advertised that before a date, a teen girl should choose an appropriate flavor because that would be her date's first taste when his lips kissed hers
Natural makeup companies have made progress in creating lip gloss with mainly natural ingredients except for preservatives.
Lip Gloss
Moisturizing Lip Gloss, Glamour Gloss, Shiny Lip Gloss, Glow in the Dark Lip Gloss, Lip Gloss Collections
Zhejiang Zhongyimei Industry Co., Ltd , https://www.queeeny.com