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| Assay Type | Sandwich-ELISA |
| Analyte | AAV5 |
| Format | 96T |
| Regulatory Status | RUO |
| Sensitivity | < 1.70E+07 capsids/mL |
| Standard Curve Range | 1.09E+9 capsids/mL-1.70E+07 capsids/mL |
| Assay Time | 3 hr 20 min |
| Suitable Sample Type | For the detection and quantitative determination of AAV5 capsid titration in AAV gene therapy product preparation processing. |
| Sample volume | 100 uL |
| ID | Components | Size |
| AAV05-C01 | Pre-coated Anti-AAV5 Antibody Microplate | 1 plate |
| AAV05-C02 | AAV5 Standard | 8.7E+10capsids |
| AAV05-C03 | Biotin-Anti-AAV5 Antibody | 20 μg |
| AAV05-C04 | Streptavidin-HRP | 50 μL |
| AAV05-C05 | 10xWashing Buffer | 50 mL |
| AAV05-C06 | 2xDilution Buffer | 50 mL |
| AAV05-C07 | Substrate Solution | 12 mL |
| AAV05-C08 | Stop Solution | 7 mL |
AAV5 Titration ELISA Kit was developed for the detection and quantitative determination of AAV5 capsid titration in AAV gene therapy product preparation processing.
It is for research use only.
The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
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 AAV5 capsid as standard. The samples and standard are diluted by Dilution Buffer.
c) Wash the plate and add the Biotin-Anti-AAV5 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.
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.
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision, Intra-Assay Precision CV＜10%.
Three samples of known concentration were tested in three separate assays to assess inter-assay precision, Inter-Assay Precision CV<10%.
Three AAV5 capsid with different concentrations were tested to calculate the recovery rate.
The specific binding of AAV-A005 to AAV1/2/3/8/9/dj serotypes was detected. The results showed that AAV-A005 only binds specifically to AAV5.
The specific binding of AAV-A005 to native/denatured AAV5 was detected. The results showed that AAV-A005 only binds specifically to native AAV5. (Denatured AAV5: 95℃ for 10min)
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