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- 详细信息
- 文献和实验
- 技术资料
- 样本:
血清、血浆、尿液、组织、上清液
- 库存:
60
- 标记物:
详见官方网站
- 适应物种:
Human
- 应用:
ELISA
- 检测方法:
酶联免疫方法
- 检测范围:
详见说明书
- 供应商:
钰博生物
- 规格:
96 tests
| For quantitative detection in human samples. | (no photo available) | 96 tests | $975 | KT-65703 |
COMPONENTS
| Reagents | Quantity |
| Pre-coated, ready to use 96-well strip plate | 1 |
| Calibrator | 2 |
| Calibrator Diluent | 1 × 20 mL |
| Detection Reagent A | 1 × 120 uL |
| Detection Reagent B | 1 × 120 uL |
| Assay Diluent A | 1 × 12 mL |
| Assay Diluent B | 1 × 12 mL |
| TMB Substrate | 1 × 9 mL |
| Stop Solution | 1 × 6 mL |
| Wash Buffer (30X concentrate) | 1 × 20 mL |
| Plate sealer for 96 wells | 4 |
- Microplate reader with 450 ± 10 nm filter.
- Precision single or multi-channel pipettes and disposable tips.
- Eppendorf Tubes for diluting samples.
- De-ionized or distilled water.
- Absorbent paper for blotting the microtiter plate.
- Container for Wash Solution. STORAGE All the reagents should be kept according to the labels on vials. The Calibrator, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon being received while the others should be at 4°C. The unused strips should be kept in a sealed bag with the desiccant provided to minimize exposure to damp air. Opened test kits will remain stable for 1 month, provided it is stored as prescribed above.
Plasma
Collect plasma using 3.8% sodium citrate (sodium citrate:blood = 1:9) as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g at 4°C within 30 minutes of collection. Remove plasma and assay immediately or store
samples in aliquot at -20°C or -80°C for later use. Avoid repeated freeze/thaw cycles. Note:
- Samples to be used within 5 days may be stored at 4°C, otherwise samples must be stored at -20°C (≤1 month) or -80°C (≤2 months) to avoid loss of bioactivity and contamination.
- When performing the assay slowly bring samples to room temperature. 3. Sample hemolysis will influence the result, so hemolytic specimen can not be detected.
Bring all kit components and samples to room temperature (18-25°C) before use.
Detection Reagent A and B
Briefly spin or centrifuge the stock Detection Reagent A and Detection Reagent B before use. Dilute to the working concentration with Assay Diluent A or B, respectively (1:100).
Wash Solution
Dilute 20 mL of Wash Solution concentrate (30X) with 580 mL of de-ionized or distilled water to prepare 600 mL of Wash Solution (1X).
ASSAY PROCEDURE SUMMARY
- Prepare all reagents, samples and calibrators;
- Add 100 µL calibrator or sample to each well. Incubate 2 hours at 37°C;
- Add 100 µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
- Aspirate and wash 3 times;
- Add 100 µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
- Aspirate and wash 5 times;
- Add 90 µL Substrate Solution. Incubate 15-25 minutes at 37°C;
- Add 50 µL Stop Solution. Read at 450 nm immediately.
1. The final experimental results will be closely related to operation skills of the end users and theexperimental environments. Please make sure that sufficient samples are available.
2. Kits from different batches may be a little different in detection range, sensitivity and color developingtime. Please perform the experiment exactly according to the instruction attached in kit whileelectronic ones from our website (www.k-assay.com) is only for information.
3. Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied bymanufacturer.
4. Protect all reagents from strong light during storage and incubation. All the bottle caps of reagentsshould be covered tightly to prevent the evaporation and contamination of microorganism.
5. There may be some foggy substance in the wells when the plate is opened at the first time. It will nothave any effect on the final assay results. Do not remove microtiter plate from the storage bag untilneeded.
6. Wrong operations during the reagents preparation and loading, as well as incorrect parameter settingfor the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10 nmor less and an optical density range of 0-3 O.D. or greater at 450 ± 10 nm wavelength is acceptablefor use in absorbance measurement. Please read the instruction carefully and adjust the instrumentprior to the experiment.
7. Even the same operator might get different results in two separate experiments. In order to get betterreproducible results, the operation of every step in the assay should be controlled. Furthermore, apreliminary experiment before assay for each batch is recommended.
8. Each kit has been strictly passed Q.C. test. However, results from end users might be inconsistentwith our in-house data due to some unexpected transportation conditions or different lab equipments.
Intra-assay variance among kits from different batches might arise from above factors, too.
9. Kits from different manufacturers for the same item might produce different results, since we haven’tcompared our products with other manufacturers.
10. The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, andclothing protection when using this material.
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文献和实验有替代前者的趋势。由于ABS-ELISA较普通ELISA多用了两种试剂,增加了操作步骤,在临床检验中ABS-ELISA应用不多。科研项目中检测微量的成分如细胞因子常采用本法。晶美分装ELISA KIT采用的方法:1, TORCH及传染病试剂盒(间接法),见2.2.32, TORCH-IgM捕获法特色:包被抗体,标记抗原原理:3, 细胞因子试剂盒采用的方法路线(ABC-ELISA)原理产品特色:采用ABC法,灵敏度更高,特异性更强。生物素抗体和酶联物是浓缩的,使用前需用相应的缓冲液稀释。酶联物可以通用
的原理用于分子诊断方面:TrimGen公司的KRAS Mutation Detection Kit在96孔板里一次能实现7/8个KRAS突变位点的检测。 另外基于相同原理的技术还有PPT-ELISA。这是用来筛截短突变的,相关文章见:1,Rapid screen for truncating ATM mutations by PTT-ELISA . 2,An ELISA-based high throughput protein truncation test
时对一个样本中的75个目的蛋白进行精确定量。2、样本需要量小。仅需要50μL溶液就可进行一次检测。3、更高的灵敏度。分析灵敏度高达2.8pg/mL。4、更高的灵敏度、更宽的检测范围和更好的重复性。CBA技术使蛋白定量的分析灵敏度高达pg/ml级别,检测范围达0-5000pg/ml,所有分析只需一组标准曲线。根据抗体的荧光强度对目的蛋白定量,排除了ELISA反应中酶联放大产生的假阳性。5、检测对象的灵活组合—BD CBA Flex Set。CBA kit是同时检测试剂盒配好的6个指标,形式比较固定
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