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: 10200000 capsids/mL - 650000000 capsids/mL

Minimum Detection Limit: 10200000 capsids/mL

Application: ELISA

Purpose: This kit uses sandwich ELISA to determine the titer of AAV9 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: 10000000 capsids/mL

Components:

• Pre-coated 96-well plate
• Reference Standard
• Detection Antibody
• HRP Conjugate
• Assay Buffer (10 x concentrate)
• Wash Buffer (20 x concentrate)
• 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. Add 100μL standard or sample to each well. Incubate 1 hour at 37 °C,
3. Aspirate and wash with 300 µL 1x Wash buffer 4 times,
4. Add 100μL Detection Antibody. Incubate 1 hour at 37 °C,
5. Aspirate and wash with 300 µL 1x Wash buffer 4 times,
6. Add 100μL Streptavidin-HRP conjugate. Incubate 1 hour at 37 °C,
7. Aspirate and wash with 300 µL 1x Wash buffer 4 times,
8. Add 100 μL TMB Substrate. Incubate 10 minutes in the dark at 37 °C,
9. Add 50μL Stop Solution. Read at 450nm immediately.

Reagent Preparation:

1. Prepare 1X Wash Buffer (Dilute 20X Wash Buffer with distilled H2O)
2. Prepare 1X Assay Buffer (Dilute 10X Assay Buffer with distilled H2O)
3. Prepare 1X Detection Antibody (Dilute the detection antibody (500X) with 1X Assay Buffer)
4. Preparation of AAV9 capsid standards: Add 1mL of distilled H2O to one vial of standard. Dissolve at room temperature for 20 min. Centrifuge at 1000 rpm for 30 sec. The titer of the reconstituted standard is 6.50E+08 capsids/mL. Two-fold serial dilution of the AAV9 standard with 1X Assay Buffer for the titration curve in duplicates is recommended.
5. Sample preparation: The supernatant of cell culture could be used directly or after dilution with cell lysis buffer (not provided). Cells should be treated with lysis buffer (not provided) and diluted with 1X assay buffer for sample preparation. Dilute the sample with 1X Assay Buffer to make the capsid titer quantification fall within the linear range.
   6.50E+08 capsids/mL, 3.25E+08 capsids/mL, 1.63E+08 capsids/mL, 8.13E+07 capsids/mL, 4.06E+07 capsids/mL, 2.03E+07 capsids/mL, 1.02E+07 capsids/mL.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof 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 therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Assay Precision: CV < 5% indicates high accuracy. CV should be < 10% for each standard and sample. Retest any samples with CV > 10%.

Restrictions: For Research Use only

Handling

Storage: 4 °C/-20 °C

Storage Comment: Store the standard and detection antibody at -20 °C and the other kit components at 2-8 °C. The kit is valid for 6 months from the production date.

Expiry Date: 6 months

Target Details for AAV9

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

Alternative Name: AAV9 (AAV9 Products)