AAV9 ELISA Kit

Catalog No. ABIN7448144

Product Details AAV9 ELISA Kit

Target: AAV9 (Adeno-Associated Virus 9 (AAV9))

Reactivity: Adeno-Associated Virus 9 (AAV-9)

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 20300000 capsids/mL - 1300000000 capsids/mL

Minimum Detection Limit: 20300000 capsids/mL

Application: ELISA

Purpose: This kit uses sandwich ELISA to determine the titer of AAV capsids in the test sample.

Sample Type: Cell Culture Supernatant, Cell Lysate

Analytical Method: Quantitative

Specificity: Intact AAV wild type virions, AAV recombinant virions, Assembled AAV virions, Intact empty AAV capsids

Sensitivity: 10200000 capsids/mL

Components:

• Pre-coated 96-well plate
• Reference Standard
• Detection Antibody (20x)
• HRP Conjugate
• Assay Buffer (20x)
• TMB Substrate
• Stop Solution
• Instruction manual

Application Details

Application Notes: Empty AAV9 capsids lacking the transgene of interest cause unwanted immune response without providing the therapeutic benefit of the transgene. Alongside DNA and infectious titer, determination of the capsid titer is crucial for validating the efficacy and safety of your AAV gene therapy. Our titration kit detects full and empty AAV9 capsids. Note: User has the option of a rapid protocol (2 hours) for fast turnaround or a standard protocol (6 hours) for greater sensitivity.

Sample Volume: 100 µL

Assay Time: 2 h

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards,
2. Wash plate with 1X Assay buffer
3. Add 100μL standard or sample to each well. Incubate 1 hour at 37 °C,
4. Aspirate and wash with 300 µL 1x Assay buffer 3 times,
5. Add 100μL Detection Antibody. Incubate 1 hour at 37 °C,
6. Aspirate and wash with 300 µL 1x Assay buffer 3 times,
7. Add 100μL Streptavidin-HRP conjugate. Incubate 1 hour at 37 °C,
8. Aspirate and wash with 300 µL 1x Assay buffer 4 times,
9. Add 100 μL TMB Substrate. Incubate 10 minutes in the dark at 37 °C,
10. Add 50μL Stop Solution. Read at 450nm immediately.

Reagent Preparation:

1. Prepare 1X Assay Buffer (Dilute 20X Assay Buffer with distilled H2O)
2. Prepare 1X Detection Antibody (Dilute the detection antibody (20X) with 1X Assay Buffer)
3. Preparation of AAV capsid standards: Add 700μL of distilled H2O to one vial of standard. Dissolve at room temperature for 10-20 min, mix gently and avoid vortexing. Two-fold serial dilution of the AAV8 standard with 1X Assay Buffer for the titration curve in duplicates is recommended.
4. Sample preparation: Dilute the sample with 1X Assay Buffer to make the capsid titer quantification fall within the linear range.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturation may occur in these samples, leading to false results. Samples should therefore be stored for a short period at 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thaw cycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged to remove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determine compatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibility of causing a deviation due to the introduced chemical substance. The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in the range of the standard curve, the optimal sample dilution for the particular experiment has to be determined.

Assay Precision: Retest any samples with CV > 10%.

Restrictions: For Research Use only

Handling

Storage: 4 °C/-20 °C

Storage Comment: Store kit components at 2-8°C. The unopened kit is valid for 12 months from the production date. Reconstituted AAV standard stable at 2-8°C for 2 weeks. -20°C or below for long-term storage and avoid repeated freeze-thaw cycles.

Expiry Date: 6 months

Target Details for AAV9

Target: AAV9 (Adeno-Associated Virus 9 (AAV9))

Alternative Name: AAV9 (AAV9 Products)