膜杰作
Star Staining
组分（Materials Provided）
| ID | Components | Size |
| AAV09H-C01 | Pre-coated Anti-AAV9 Antibody Microplate | 1 plate |
| AAV09H-C02 | AAV9 Standard | 3.80E+11 capsids |
| AAV09H-C03 | HRP-Anti-AAV9 Antibody | 20 μg |
| AAV09H-C04 | 10×Washing Buffer | 50 mL |
| AAV09H-C05 | 2×Dilution Buffer | 50 mL |
| AAV09H-C06 | Substrate Solution | 12 mL |
| AAV09H-C07 | Stop Solution | 7 mL |
背景（Background）
Adeno-associated virus (AAV) has become one of the most important gene vectors in the field of gene therapy due to its long-term expression, low toxicity, low immunogenicity, and high tissue specificity.Most successful AAV gene therapies for preclinical and clinical studies are limited to natural serotypes, but the presence of neutralizing antibodies against AAV remains a significant barrier to systemic delivery.
应用说明（Application）
AAV9 Titration ELISA Kit was developed for the detection and quantitative determination of AAV9 capsid titration in AAV gene therapy 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）
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 AAV9 capsid. The kit consists of Pre-coated Anti-AAV9 Antibody Microplate and AAV9 Standard,HRP-Anti-AAV9 Antibody and buffers.
Your experiment will include 5 simple steps:
a) Bring all reagents and samples to room temperature (20℃-25℃) before use.
b) Add your sample to the plate, take the AAV9 capsid as Control sample. The samples and Control sample are diluted by Dilution Buffer.
c) Add the HRP-Anti-AAV9 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 prior to statistical analysis. The OD Value reflects the amount of protein 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 AAV9 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 AAV9 with different concentrations were tested to calculate the recovery rate.
专属性（Specificity）
The specific binding of AAV-A009H to AAV1/2/3/5/6/8/9/dj serotypes was detected. The results showed that AAV-A009H only binds specifically to AAV9.
The specific binding of AAV-A009H to native/denatured AAV9 was detected. The results showed that AAV-A009H only binds specifically to native AAV9. (Denatured AAV9: 95℃ for 5min)
背景（Background）：AAV9
The adeno-associated virus (AAV) is a small (25 nm), non-enveloped virus of the parvoviridae family, including 12 different AAV serotypes. In the parvoviridae family it belongs to the genus dependoparvovirus, because it needs the presence of a helper virus for replication and assembly. The icosahedral AAV capsid composed of the capsid proteins VP1, VP2 and VP3 contains a linear, single-stranded DNA genome of 4.7 kb.
