膜杰作
Star Staining
产品描述（Product Details）
| Assay Type | Sandwich-ELISA |
| Analyte | TPO |
| Format | 96T |
| Reactivity | Human |
| Regulatory Status | RUO |
| Sensitivity | ＜12.5 pg/mL |
| Standard Curve Range | 12.5 pg/mL-400 pg/mL |
| Assay Time | 3 hr 20 min |
| Suitable Sample Type | For the detection and quantitative determination of TPO in human serum and cell culture supernates. |
| Sample volume | 100 uL |
背景（Background）
Human TPO Kit is based on ELISA sandwich method and designed to measure human TPO levels in cell culture supernates, serum, and plasma. It contains recombinant human TPO and a pair of antibodies against the recombinant human TPO, which are provided by ACROBiosystems. Results are obtained by four parameter logistic curve that were parallel to the standard curves obtained. The verification results indicate that this kit can be used for the quantitative determination of GMP human TPO (ACROBiosystems, cat#GMP-THNH25) concentrations. The specificity has been verified.
应用说明（Application）
The kit is developed for the detection and quantitative determination of TPO in human serum and cell culture supernates.
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.
组分（Materials Provided）
| ID | Components | Size |
| RES034-C01 | Pre-coated Anti-TPO Antibody Microplate | 1 plate |
| RES034-C02 | Human TPO Standard | 20 μg |
| RES034-C03 | Biotin-Anti-TPO Antibody | 20 μg |
| RES034-C04 | Streptavidin-HRP | 50 μL |
| RES034-C05 | 10×Washing Buffer | 50 mL |
| RES034-C06 | 2×Dilution Buffer | 50 mL |
| RES034-C07 | Substrate Solution | 12 mL |
| RES034-C08 | Stop Solution | 7 mL |
原理（Assay Principles）
This assay kit employs a standard sandwich-ELISA format, providing a rapid detection of Human TPO. The kit consists of Pre-coated Anti-TPO Antibody Microplate and Human TPO Standard and Biotin-Anti-TPO 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 TPO as standard. The samples and standard are diluted by Dilution Buffer.
c) Wash the plate and add the Biotin-Anti-TPO 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 TPO 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 TPO, and the T cell culture supernatant without human TPO was used as background to calculate the recovery rate. The range of the recovery rate is 94.7-108.9%, and the average recovery is 102.1%.
背景（Background）：TPO
Thrombopoietin (TPO) is a 332 amino acid glycoprotein made primarily in the liver and act as the major physiological regulator of platelet production. TPO binds the TPO receptor, activates JAK and STAT pathways, thus stimulating megakaryocyte growth and platelet production. Additionally, further investigation uncovered that thrombopoietin is a critical cytokine promoting hematopoietic rebound after myeloablation and its transcripts are expressed by multiple cellular sources.
