膜杰作
Star Staining
产品描述（Product Details）
| Assay Type | Sandwich-ELISA |
| Analyte | IFN-γ |
| Format | 96T |
| Reactivity | Human |
| Regulatory Status | RUO |
| Sensitivity | < 3.125 pg/mL |
| Standard Curve Range | 3.125-200 pg/mL |
| Assay Time | 2 hr 50 min |
| Suitable Sample Type | For the quantitative determination of human IFN-γ in Cell Culture Supernatants, Plasma, Serum. |
| Sample volume | 100 uL |
组分（Materials Provided）
| ID | Components | Size |
| RES028-C01 | Pre-coated Anti-IFN-γ Antibody Microplate | 1 plate |
| RES028-C02 | Human IFN-γ Standard | 20 μg |
| RES028-C03 | Biotin-Anti-IFN-γ Antibody | 50 μL |
| RES028-C04 | Streptavidin-HRP | 50 μL |
| RES028-C05 | 10xWashing Buffer | 50 mL |
| RES028-C06 | 2xDilution Buffer | 50 mL |
| RES028-C07 | Substrate Solution | 12 mL |
| RES028-C08 | Stop Solution | 7 mL |
背景（Background）
Human Interferon-γ (IFN-γ) ELISA Kit is based on ELISA sandwich method and designed to measure human IFN-γ levels in cell culture supernates, serum, and plasma. It contains recombinant human IFN-γ and a pair of antibodies against the recombinant factor, 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 Interferon-γ (IFN-γ) (ACROBiosystems, cat#GMP-IFGH24) concentrations. The specificity has been verified.
应用说明（Application）
The Human Interferon-γ (IFN-γ) (Residue Testing)was developed for the detection and quantitative determination of GMP human IFN-γ 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）
Unopened kit should be stored at 2°C -8°C upon receiving.
Find the expiration date on the outside packaging and do not use reagents past their expiration date.
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 sandwich-ELISA format, providing a rapid detection of Human IFN-γ. The kit consists of Pre-coated Anti-IFN-γ Antibody Microplate and Human IFN-γ Standard and Biotin-Anti-IFN-γ 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 IFN-γ as standard. The samples and standard are diluted by Dilution Buffer.
c) Wash the plate and add the Biotin-Anti-IFN-γ 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 IFN-γ, 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 twenty times on one plate to assess intra-assay precision, Intra-Assay Precision CV＜10%.
批间差异（Inter-Assay Statistics）
Three samples of known concentration were tested in three separate assays to assess inter-assay precision, Inter-Assay Precision CV<10%.
回收率（Recovery）
Three parts of blank serum were added with different concentrations of human IFN-γ and the serum without human IFN-γ was used as background to calculate the recovery rate. The range of the recovery rate is 89.2-116.0%, and the average recovery is 98.3%.
背景（Background）：IFN-gamma
Interferon gamma, Interferon-γ
Interferon-gamma (IFN-γ/IFNG) is a dimerized soluble cytokine that is the only member of the type II class of interferon. This interferon was originally called macrophage-activating factor, a term now used to describe a larger family of proteins to which IFN-γ belongs. IFN-gamma has been used in a wide variety of clinical indications. Interferon-gamma (IFNgamma) is a central regulator of the immune response and signals via the Janus Activated Kinase (JAK)-Signal Transducer and Activator of Transcription (STAT) pathway. Interferon gamma has broader roles in activation of innate and adaptive immune responses to viruses and tumors, in part through upregulating transcription of genes involved in cell cycle regulation, apoptosis, and antigen processing/presentation. Despite this, rodent and human trophoblast cells show dampened responses to IFNG that reflect the resistance of these cells to IFNG-mediated activation of major histocompatibility complex (MHC) class II transplantation antigen expression.
Cytokine storm syndromes is an activation cascade of pro-inflammatory cytokines，such as TNF-α，IL-1，IL-6，IL-12，IFN-α，IFN-β，IFN-γ，MCP-1 and IL-8,due to unregulated host immune response system to different stimuli such as infections, malignancy, rheumatoid disorders, drugs, and so on. Interferon-γ (IFN-γ), one of the four subclasses ofinterferons, and is a multipotent protein, acting on many cell types by inducing or inhibiting many cellular functions through direct effects on gene expression.
