0
Boster's ELISA Kit is a competitive assay that can be used for the quantification of Free Nitrotyrosine in plasma, serum, cell lysates, urine, and other sample matrices. The ELISA utilizes an Nitrotyrosine-coated plate and an HRP-conjugated antibody for detection which allows for an assay range of 62.5 to 8000nM Free Nitrotyrosine, with a sensitivity of 50nM. The other highlights of this kit are a quick incubation time of 60 minutes, stable reagents, and an easy to use protocol.
| Product Name | Nitrotyrosine ELISA Kit |
|---|---|
| SKU/Catalog Number | EK7116 |
| Description | Colorimetric detection of Nitrotyrosine. 96wells/kit, with removable strips. |
| Cite This Product | Nitrotyrosine ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK7116) |
| Validated Species | All Animals |
| Application | ELISA *Our Boster Guarantee covers the use of this product in the above tested applications. |
| Cross Reactivity | There is no detectable cross-reactivity. |
| Pack Size | 96wells/kit, with removable strips. |
| Sensitivity | 50 nM *Sensitivity, or Lower Limit of Detection (LLD), is the minimum level of target protein the ELISA assay can detect. We measure 20 blank wells and if the O.D. value is 2 standard deviations higher than the blanks' average O.D. the sample can be deemed positive. |
|---|---|
| Assay Range | 62.5 - 8000 nM *This assay range is determined using common samples. For samples with low target protein concentrations, users can adjust the standard curve to extend the lower limit of assay range. |
| Sample Type | Cell lysates, Plasma, Serum, Urine *The above listed samples are the ones valided with the assay. If you do not see your sample of interest listed, as long as there is enough level of target protein present in the sample, this Picokine? ELISA kit should detect it. **For protocol and tips regarding preparing your sample of interest, please check our ELISA sample preparation guide. |
| Storage | Store at 4°C. |
| Description | Quantity |
| Nitrosylated BSA Coated Plate | 1 Plate |
| Nitrotyrosine Standard | 1 vial/110?l |
| Nitrotyrosine HRP Conjugated Monoclonal Antibody | 1 vial/75?l |
| Sample and Standard Diluent (Red) | 1 vial/50mL |
| Nitrotyrosine Antibody Diluent (Blue) | 1 vial/13mL |
| Wash Buffer Concentrate (10X) | 1 vial/ 50mL |
| TMB Substrate | 1 vial/ 13 ml |
| Stop Solution | 1 vial/ 13 ml |
| Plate Cover | 2 |
1. A plate reader capable of measuring absorbance at 450 nm.
2. Adjustable pipettes and a repeat pipettor.
3. Deionized or distilled water
4. Materials used for Sample Preparation
Nitrotyrosine has been identified as a marker of inflammation and NO production. Nitrotyrosine is formed in presence of the active metabolite NO. Various pathways including the formation of peroxinitrite lead to nitrotyrosine production. Since nitrotyrosine is a stable end product of peroxynitrite oxidation, assessment of its plasma concentration may be useful as a marker of NO-dependent damage in vivo. Since NOX is only an indicator for enhanced NO production, protein associated nitrotyrosine might be a more suitable marker for damage induced by reactive nitrogen intermediates derived from NO. Furthermore, most proteins have a longer half life in the circulation than NOX levels. The presence of nitrotyrosine has been detected in various inflammatory processes including atherosclerotic plaques, celiac disease, rheumatoid arthritis, chronic renal failure and septic shock. In normal plasma low, undetectable, levels of nitrotyrosine are present. Nitrosylation of the amino acid tyrosine occurs both for free tyrosine and for protein bound tyrosine.
Click the images to enlarge.
Typical Standard Curve for the Nitrotyrosine ELISA Kit (Enzyme-Linked Immunosorbent Assay)–EK7116 Assay Type: Competitive ELISA. Detection Method: Colorimetric Assay. Assay Range: 62.5 – 8000 nM.
Diagram of the Competitive ELISA
Diagram of the Preparation of Nitrotyrosine Standards
Diagram of the Triplicate Sample Plate Format
Intra-Assay Precision: Three samples of known concentration were assayed thirty times on one plate. The intra-assay coefficient of variation of the Nitrotyrosine ELISA has been determined to be <10%.
Inter-Assay Precision: Three samples of known concentration were assayed thirty times in three individual assays. The inter-assay coefficient of variation of the Nitrotyrosine ELISA has been determined to be <15%.
1. Prepare standard and samples in the Sample and Standard Diluent.
2. Add 50 ?L of prepared standards and samples in triplicate to appropriate wells.
3. Add 50 ?L of the diluted antibody preparation to the appropriate wells.
4. Cover plate with Plate Cover and incubate at room temperature (20-25°C) for 1 hour.
5. Wash plate 4 times with 1X Wash Buffer.
6. Add 100 ?L of TMB Substrate to each well.
7. Cover plate and develop the plate in the dark at room temperature for 30 minutes.
8. Add 100 ?L of Stop Solution to each well.
9. Measure absorbance on a plate reader at 450 nm.
10. Plot the standard curve and calculate sample concentrations.
温馨提示:因厂家更改产品包装、产地或者更换随机附件等没有任何提前通知，且每位咨询者购买情况、提问时间等不同，为此以下回复仅对提问者3天内有效，其他网友仅供参考！若由此给您带来不便请多多谅解，谢谢！
服务热线
0771-3293894
新浪微博
腾讯微博
QQ空间
客服
