膜杰作
Star Staining
产品描述（Product Details）
| Assay Type | Sandwich-ELISA |
| Analyte | IL-10 |
| Format | 96T |
| Reactivity | Human |
| Regulatory Status | RUO |
| Sensitivity | ＜6.25 pg/mL |
| Standard Curve Range | 6.25 pg/mL-200 pg/mL |
| Assay Time | 2 hr 50 min |
| Suitable Sample Type | For detection and quantitative determination of GMP human IL-10 in samples from CAR-T product preparation processing. |
| Sample volume | 100 uL |
背景（Background）
IL-10 is an anti-inflammatory cytokine. It maintains the balance of the immune response, allowing the clearance of infection while minimizing damage to the host by inhibits the activity of Th1 cells, NK cells, and macrophages. The receptor for IL10 is a heterotetramer complex comprising two IL10Rα molecules and two IL10Rβ molecules .
应用说明（Application）
Human Interleukin-10 (IL-10) ELISA Kit (Residue Testing) was developed for the detection and quantitative determination of GMP human IL-10 in samples from CAR-T product preparation processing.
It is for research use only.
重构方法（Reconstitution）
Please see Certificate of Analysis for details of reconstitution instruction and specific concentration.
存储（Storage）
The unopened kit is stable for 12 months from the date of manufacture if stored at 2°C to 8°C.
The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
组分（Materials Provided）
| ID | Components | Size |
| CRS008-C01 | Pre-coated Anti-IL-10 Antibody Microplate | 1 plate |
| CRS008-C02 | Human IL-10 Standard | 20 μg |
| CRS008-C03 | Biotin-Anti-IL-10 Antibody | 50 μL |
| CRS008-C04 | Streptavidin-HRP | 50 μL |
| CRS008-C05 | 10×Washing Buffer | 50 mL |
| CRS008-C06 | 2×Dilution Buffer | 50 mL |
| CRS008-C07 | Substrate Solution | 12 mL |
| CRS008-C08 | Stop Solution | 7 mL |
原理（Assay Principles）
This assay kit employs a standard sandwich-ELISA format, providing a rapid detection of Human IL-10. The kit consists of Pre-coated Anti-IL-10 Antibody Microplate and Human IL-10 Standard and Biotin-Anti-IL-10 Antibody and Streptavidin-HRP and buffers.
Your experiment will include 6 simple steps:
a) Bring all reagents to room temperature(20℃-25℃) before use.
b) Add your sample to the plate and take the Human IL-10 as standard. The samples and standard are diluted by Dilution Buffer.
c) Wash the plate and add the Biotin-Anti-IL-10 Antibody diluted by Dilution Buffer to the plate.
d) Wash the plate and add the Streptavidin-HRP diluted by Dilution Buffer to the plate.
e) Wash the plate and add TMB.
f) Stop the substrate reaction by adding diluted acid. Absorbance (OD) is calculated by the absorbance at 450 nm minus the absorbance at 630 nm to remove background disturbance before statistical analysis. The OD Value reflects the amount of bound protein.
典型数据-Typical Data Please refer to DS document for the assay protocol.
For each experiment, a standard curve needs to be set for each micro-plate, and the specific OD value may vary depending on different laboratories, testers, or equipments. The following example data is for reference only.
验证（Validation）
稀释线性（Dilution Linearity）
To assess the linearity of the assay, samples spiked with high concentrations of human IL-10 were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.
批内差异（Intra-Assay Statistics）
Three samples of known concentration were tested ten times on one plate to assess intra-assay precision.
批间差异（Inter-Assay Statistics）
Three samples of known concentration were tested in three separate assays to assess inter-assay precision.
回收率（Recovery）
Five parts of blank T cell culture supernatant were added with different concentrations of human IL-10, and the T cell culture supernatant without human IL-10 was used as background to calculate the recovery rate. The range of the recovery rate is 86.6-95.0%, and the average recovery is 91.9%.
