上海逸峰生物科技有限公司
2011/12/31 9:06:02FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
Porcine Cortisol ELISA Kit instruction
Kit name
Porcine Cortisol ELISA Kit
Intended use
The kit is used to assay the content of Porcine Cortisol in Porcine serum,blood plasma and other related tissue liquid.
Test principle
The kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) to assay the level of Porcine Cortisol in samples. Add Porcine Cortisol to pre-coated Porcine Cortisol monoclonal antibody microelisa well, incubation; washing. Add HRP tagged Porcine Cortisol antibodies. After another incubation and washing, remove the unbound enzyme, add Chromogen Solution A and B, the color of the liquid change into blue, and the color finally become yellow at the effect of acid. The depth of the color is positively correlated with concentration of the Porcine Cortisol in samples.
Materials supplied
1 | Microelisa Stripplate | 12well×8strips | 7 | Chromogen Solution A | 6mL |
2 | Standard:80ng/mL | 0.6mL | 8 | Chromogen Solution B | 6mL |
3 | 20×wash solution | 25mL | 9 | Stop Solution | 6mL |
4 | Standard diluent | 6mL | 10 | Instruction | 1 |
5 | Sample diluent | 6mL | 11 | Closure plate membrane | 2 |
6 | HRP-Conjugate Reagent | 6mL | 12 | Sealed bags | 1 |
Note: Standard was diluent with Standard diluent followed by: 80、40、20、10、5、2.5ng/mL
Materials required but not supplied
1.
2. Standard microplate reader
3. Precision pipettes and Disposable pipette tips
4. Distilled water
5. Disposable tubes for sample dilution
6. Absorbent paper
Assay procedure
1. Prepare: The kit takeing out from the environment of 2
2. Diluent: Diluent the 20×wash solution.
3. Add standard and Sample: Set Standard wells, testing sample wells and blank wells. Add Diluted standard 50μl to standard well; Add Sample dilution 40μl to testing sample well which on Assay plate, then add testing sample 10μl (sample final dilute degree is 5 times), blank well doesn’t add anyting.
4. Incubation: Incubate 30 minutes at
5.
6. Add HRP-conjugate reagent: Add HRP-conjugate reagent 50μl to each well, except the blank well. Mixing gently shaking, incubated 30 minutes at
7. Repeat step4.
8. Repeat step5
9. Add chromogen solution A and B: Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix, incubate for 15 min at
10. Add Stop Solution: Add Stop Solution 50μl to each well, Stop the reaction(the blue color change to yellow immediay).
11. Take blank well as zero, measure the optical densit (OD) at 450 nm after adding Stop Solution and within 15 min.
12. According to standard concentration and the corresponding OD values calculated standard curve linear regression equation, then the OD values according to the sample on the regression equation to calculate the corresponding sample concentration. It should be remembered that the sample has been diluted and its actual concentration should be multiplied by the total dilution.
Specimen requirements
1. Can’t detect the samples which contain NaN3, because NaN3 inhibits HRP activity of the horseradish peroxidase.
2. Extract as soon as possible after Specimen collection, Extracted according to the relevant literature, and should be experiment as soon as possible after the extraction. If it can not be tested immediay, specimen can be kept in
3. The samples shoule be centrifugated dequay and no hemolysis or granule was allowed.
Important notes
1. The operation should be carried out in strict accordance with instructions and test results must be based on microplate reader to determine readings shall prevail.
2. If the microelisa stripplate has not used up after open, it should be stored in the sealed bag.
3. Recommended that all standard materials, test samples are doing double to minish the Experimental error.
4. Please multiply total dilution times when calculation. 5 times is the best dilute time according to this ELISA Kit design.
5. If the testing material content in the sample is excessively high, please use Special dilution to dilute certain multiple, then assay.
6. If the color too shallow, It may be appropriate to extend the substrate incubation time.
7. Add Sample with sampler each step and proofread its accuracy frequently to avoid the experimental error. In order to avoid cross-contamination, avoid to reusing the suction head and closure plate membrane.
8. Use the kit in validity, not mix the reagents of different batches.
9. Chromogen Solution B is light-sensitive, avoid prolonged exposure to light,
Summary procedures
Preparing reagents, samples and standard
Add prepared sample and standard, incubated 30 minutes at
Plate washed four times, adding HRP-Conjugate Reagent incubated 30 minutes at
Add stop solution
Measure within 15min
Calculation
Assay range:2.5-80ng/mL
Package size: 96 determinations
Storage: 2
validity: six months.
上海逸峰生物科技有限公司代理不同品牌价格档次的ELISA试剂盒。数万种抗体产品等, 品种多,质量好,灵敏度高,价格实惠,并且还提供免费代检测服务。
本公司的更多产品,请点击公司:http://www.yfswbio.com/
订货:
网 站:http://www.yfswbio.com :yfswbio.cn