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- 详细信息
- 文献和实验
- 技术资料
- 库存:
50
- 英文名:
TGFBI/beta IG-H3 ELISA
- 保质期:
12个月
- 供应商:
钰博生物
- 保存条件:
-20°C
- 规格:
1 kit
| For quantitative detection of human TGFBI in cell culture supernates, serum, and plasma (heparin, EDTA). Assay Range: 62.5 - 4,000 pg/mL. | (no photo available) | 1 kit | $467 | KT-1474 |
| 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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文献和实验一、ELISA的原理 ELISA的基础是抗原或抗体的固相化及抗原或抗体的酶标记。结合在固相载体表面的抗原或抗体仍保持其免疫学活性,酶标记的抗原或抗体既保留其免疫学活性,又保留酶的活性。在测定时,受检标本(测定其中的抗体或抗原)与固相载体表面的抗原或抗体起反应。用洗涤的方法使固相载体上形成的抗原抗体复合物与液体中的其他物质分开。再加入酶标记的抗原或抗体,也通过反应而结合在固相载体上。此时固相上的酶量与标本中受检物质的量呈一定的比例。加入酶反应的底物后,底物被酶催化成为有色产物,产物的量与标本中
抗体的结构 抗体是能与抗原特异性结合的免疫球蛋白(immunoglobulin,Ig)。Ig 分五类,即 IgG、IgA、IgM、IgD 和 IgE。与免疫测定有关的 Ig 主要为 IgG 和 IgM。Ig 由两个轻链(L)和两个重链(H)的单体组成。Ig 的轻链是相同的,有 κ(kappa)和 λ(Lambda)两种型别。五类 Ig 的重链结构不同,这决定了它们的抗原性也不同。IgG 和 IgM 的重链分别称为 γ(gamma)链和 μ(mu )链。 重链和轻链的 N 端的氨基酸排列顺序
一、ELISA的原理ELISA的基础是抗原或抗体的固相化及抗原或抗体的酶标记。结合在固相载体表面的抗原或抗体仍保持其免疫学活性,酶标记的抗原或抗体既保留其免疫学活性,又保留酶的活性。在测定时,受检标本(测定其中的抗体或抗原)与固相载体表面的抗原或抗体起反应。用洗涤的方法使固相载体上形成的抗原抗体复合物与液体中的其他物质分开。再加入酶标记的抗原或抗体,也通过反应而结合在固相载体上。此时固相上的酶量与标本中受检物质的量呈一定的比例。加入酶反应的底物后,底物被酶催化成为有色产物,产物的量与标本中受检
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