膜杰作
Star Staining
组分（Materials Provided）
| ID | Components | Size |
| RES095-C01 | Pre-coated Human CD3E & CD3G Microplate | 1 plate |
| RES095-C02 | Anti-CD3 Antibody Standard | 20 μg |
| RES095-C03 | HRP-conjugated antibody | 10 μg |
| RES095-C04 | 10×Washing Buffer | 50 mL |
| RES095-C05 | 2×Dilution Buffer | 50 mL |
| RES095-C06 | Substrate Solution | 12 mL |
| RES095-C07 | Stop Solution | 7 mL |
背景（Background）
Since the 1990s, CD3 monoclonal antibody has been used in CIK cell therapy to stimulates the proliferation and activation of T cells. Under the cooperation of other cytokines, such as IL2 and IL1a, CIK cells with rapid proliferation, high tumoricidal activity, broad tumor killing spectrum and non-MHC-restricted tumor killing characteristics are generated, which has significant effects on the treatment of cancer, chronic leukemia, liver disease and neurological diseases. Obviously, It is necessary to control the residues of raw materials in the final cell therapy product.
应用说明（Application）
The kit is developed for the detection of anti-CD3 antibody in Bioprocess manufacturing applications. It is used as a tool to aid in routine quality control of in-process streams as well as final product .
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）
1. Unopened kit should be stored at 2℃-8℃ upon receiving.
2. Find the expiration date on the outside packaging and do not use reagents past their expiration date.
3. The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
原理（Assay Principles）
This assay kit employs a standard indirect-ELISA format, providing a rapid detection of Anti-CD3 antibodies. The kit consists of Pre-coated Human CD3E & CD3G Microplate and Anti-CD3 Antibody Standard and HRP-conjugated antibody and related buffer.
Your experiment will include 5 simple steps:
a) Bring all reagents to room temperature(20℃-25℃) before use.
b) Add your sample to the plate and take the Anti-CD3 Antibody as standard. The samples and standard are diluted by Dilution Buffer.
c) Wash the plate and add the HRP-conjugated antibody diluted by Dilution Buffer to the plate.
d) Wash the plate and add TMB.
e) Stop the substrate reaction by adding diluted acid. Absorbance (OD) is calculated as 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 Antibody bound.
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 Anti-CD3 Antibody 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 Anti-CD3 Antibody, and the T cell culture supernatant without Anti-CD3 Antibody was used as background to calculate the recovery rate. The range of the recovery rate is 81.3-102.5%, and the average recovery is 89.0%.
