S100B Protein ELISA Kit
Germany DRG imported, article number: EIA5169
German DRG China exclusive agent "Shenzhen Kerunda Biological Engineering Co., Ltd." dedicated to serve you
expected usage
This kit is used for the detection of S100B concentration in human serum and plasma, only for laboratory use.
Experimental principle
This kit binds to S100B based on two antibodies, one immobilized on the coated plate and the other labeled with HRP. The test is based on a two-step method. After each plate wash, the combined and free will be separated. When the substrate liquid TMB is added, a color change will occur, the stop solution is added to terminate the reaction, and the absorption value is detected. The concentration calculation of S100B in the sample can be started from the standard. The color intensity is proportional to the concentration of S100B in the sample.
Kit composition
1. S100B standard, 6 pieces, lyophilized powder, STD0-STD5
2. Quality control, 2, freeze-dried powder, negative control and positive control
4. Enzyme conjugate, 1 branch, 1 ml, HRP-labeled anti-S100B
5. Test buffer, 1 branch, 12 ml, Tris buffer, BSA 10 g/L, stabilizer
6. Cover plate, detachable, coated with anti-S100B
7. TMB substrate solution, 1 branch, 15ml, H 2 O 2 -TMB 0.25g/L, avoid skin contact
8. Stop solution, 1 branch, 15ml, 0.15mol/L sulfuric acid, avoid skin contact
9. Wash concentrate, 50X, 1 stick, 20ml, NaCl 45g/L, Tween 20 55g/L
Required for the experiment but not provided in the kit
Deionized water
2. Automatic pipette
3. Microplate reader
Note: All reagents should be stored in the dark at 2-8 ° C, the microplate is balanced to room temperature to open the package, seal immediately after use, do not remove the adhesive strip of the unused slats
Precautions
1. The reagent contains thiomersal preservative
2. Operate and draw reagents with the highest accuracy requirements
3. All reagents have their own batch number. Do not mix substances in different batches.
4. S100B detection range is 10-5000pg/ml
5. See the label on the bottle for the specific concentration of the standard.
6. Do not use severe hemolysis samples
7. Avoid exposure of the substrate to sunlight, metal or oxidizer
Steps
Sample preparation
S100B test can use human serum or plasma, do not use hemolyzed sample, sample can be stored at 2-8 ° C for 1 day, longer shelf life can be placed at -20 ° C, avoid repeated freezing and thawing, do not place the sample at room temperature Excessive time, when the sample concentration is higher than 5ng/ml, further diluted with test buffer
Preparation of standards and controls
Standards and controls were prepared in 1 ml of deionized water and stored at 2-8 °C for 4 weeks, -20 °C for 6 months, and recommended to be stored at -20 °C. See the label for the standard concentration. Avoid repeated freezing and thawing and prolonged exposure to room temperature
Preparation of enzyme conjugates
Prepare 2 hours before use
50ul of enzyme conjugate + 1.0ml of enzyme-linked buffer (the amount of diluted enzyme conjugate is proportional to the amount of test sample), mixed for 5min to avoid bubble generation, can be stored for 3 hours at room temperature (22-28 ° C)
Washing liquid preparation
10ml of washing concentrate + 490ml of deionized water; different volumes but maintaining the same dilution ratio; at room temperature (22-28 ° C) can be saved to the expiration date shown on the label
Experimental procedure
Double testing is recommended, including standards, controls, and samples, as well as a blank control hole.
Reagent | Standard | Sample / quality control | Blank control |
Standard S0-S5 Quality control | 50ul | | |
Sample / quality control | | 50ul | |
Test buffer | 50ul | 50ul | |
|
Diluted enzyme conjugate | 100ul | 100ul | |
Incubate for 1 hour at room temperature, discard the incubation solution, and wash the plate 6 times with 300 ul of wash solution per well. |
TMB substrate solution | 100ul | 100ul | 100ul |
Incubate for 30 min at room temperature in the dark |
Stop solution | 100ul | 100ul | 100ul |
Gently shake the microplate and read on the microplate reader (450nm) |
Result calculation
1. Calculate the average OD of each standard and sample
2. Draw a most suitable standard curve (three regressions or four parameters) based on the absorbance and concentration of the standard.
3. Read sample concentration directly from the standard curve
This translation is for reference only, please refer to the original for details.
Exclusive distributor in China: Shenzhen Kerunda Bioengineering Co., Ltd.
Company address : 6th Floor, No. 10, Yanshan Road, Shekou, Nanshan District, Shenzhen. Zip code : 518067
Contact number (8 lines) Fax : +86 755 26814431
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