组分(Materials Provided)
ID | Components | Size |
CAS01-C01 | Pre-coated Anti-Cas9 Antibody Microplate | 1 plate |
CAS01-C02 | Cas9 Nuclease Standard | 20 μg |
CAS01-C03 | Biotin-Anti-Cas9 Antibody | 15 μg |
CAS01-C04 | Streptavidin-HRP | 50 μL |
CAS01-C05 | 10xWashing Buffer | 50 mL |
CAS01-C06 | 2xDilution Buffer | 50 mL |
CAS01-C07 | Substrate Solution | 12 mL |
CAS01-C08 | Stop Solution | 7 mL |
背景(Background)
CRISPR-Cas9 is the third generation of gene editing technology after ZFN and TALENs, which has the advantages of high editing efficiency, low cost, and easy operation, and has become the most mainstream gene editing method today. At present, several cell and gene therapy drugs based on CRISPR-Cas9 technology have been approved for clinical trials. In in vitro gene therapy, cells modified by the CRISPR-Cas9 system are tested for extracellular or intracellular Cas9 nuclease residues before being infused back into the body.
应用说明(Application)
resDetect™ Cas9 (CRISPR Associated Protein 9) ELISA Kit (Residue Testing) is developed for quantitative detection of Cas9 in cell Therapy.
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 sandwich-ELISA format, providing a rapid detection of Cas9 Nuclease. The kit consists of Pre-coated Anti-Cas9 Antibody Microplate and Cas9 Nuclease Standard and Biotin-Anti-Cas9 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 Cas9 Nuclease Standard. The samples and standard are diluted by Dilution Buffer.
c) Wash the plate and add the Biotin-Anti-Cas9 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 Cas9 nuclease 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)
Three Cas9 nuclease with different concentrations were tested to calculate the recovery rate.