QuickTiter™ Hepatitis B Surface Antigen (HBsAg) ELISA Kit

The QuickTiter™ Hepatitis B Surface Antigen (HBsAg) ELISA Kit is a highly sensitive sandwich ELISA assay designed for the detection and quantification of HBsAg in various biological samples, including purified virus and unpurified viral supernatants. With a sensitivity as low as 1 ng/mL, this kit provides a robust tool for studying HBV infection and monitoring antigen levels in research and diagnostic workflows.

Key Features

1. High Sensitivity and Specificity:
   • Detects HBsAg concentrations as low as 1 ng/mL with minimal cross-reactivity.
2. Quantitative Results:
   • Provides accurate measurement of HBsAg levels using a standard curve.
3. Comprehensive Kit Components:
   • Reagents sufficient for 96 wells, including standards, unknown samples, and controls.
4. Reliable Detection Methodology:
   • Utilizes a FITC-conjugated anti-HBsAg antibody and an HRP-conjugated anti-FITC antibody for enhanced specificity and signal amplification.
5. Wide Application Range:
   • Suitable for research on purified viruses and viral supernatants.

Principle of the Assay

The assay uses a sandwich ELISA technique for detecting HBsAg:

1. Capture Phase:
   • The pre-coated microplate contains anti-HBsAg antibodies that bind to HBsAg in the sample.
2. Detection Phase:
   • A FITC-conjugated anti-HBsAg antibody binds to the captured antigen-antibody complex.
   • An HRP-conjugated anti-FITC antibody binds to the FITC label, amplifying the signal.
3. Signal Development:
   • The addition of TMB substrate produces a colorimetric signal directly proportional to the HBsAg concentration in the sample.
   • The reaction is stopped, and optical density (OD) is measured at 450 nm.
4. Quantification:
   • HBsAg levels are determined using the standard curve.

Applications and Use Cases

1. Clinical and Diagnostic Research:

• HBV Infection Detection:
  • Measure HBsAg levels to assess active HBV infection.
• Disease Monitoring:
  • Track disease progression and response to therapy.

2. Vaccine and Therapeutic Development:

• Vaccine Evaluation:
  • Quantify HBsAg to study vaccine-induced immune responses.
• Drug Development:
  • Monitor antiviral drug efficacy in reducing HBsAg levels.

3. Public Health and Epidemiology:

• HBV Screening Programs:
  • Use in serological studies to evaluate HBV prevalence in populations.
• Outbreak Investigations:
  • Detect and quantify HBsAg in suspected transmission events.

4. Research in Virology:

• HBV Pathogenesis Studies:
  • Investigate the role of HBsAg in HBV infection and immune response evasion.
• Virus Production Monitoring:
  • Quantify HBsAg in viral supernatants during HBV culture.

Kit Components

• Pre-Coated 96-Well Plate: Anti-HBsAg antibody coated wells for antigen capture.
• HBsAg Standards: For generating the standard curve.
• FITC-Conjugated Anti-HBsAg Antibody: Detects the captured HBsAg.
• HRP-Conjugated Anti-FITC Antibody: Amplifies the detection signal.
• Sample Diluent: Ensures optimal sample preparation.
• Wash Buffer: Removes unbound components.
• TMB Substrate: Produces a colorimetric signal.
• Stop Solution: Stabilizes the color change for accurate OD readings.
• Instruction Manual: Step-by-step guide for assay execution.

Performance Specifications

• Sensitivity: 1 ng/mL
• Dynamic Range: Dependent on the provided standards.
• Sample Compatibility: Purified virus and unpurified viral supernatants.

Advantages

1. Ease of Use:
   • Ready-to-use reagents and pre-coated plates simplify the workflow.
2. High Accuracy:
   • Ensures reliable HBsAg quantitation with minimal variability.
3. Versatility:
   • Suitable for a wide range of HBV-related research applications.
4. Fast Results:
   • Efficient assay design provides rapid and reproducible data.

Sample Preparation Recommendations

1. Purified Virus Samples:
   • Dilute as required based on expected HBsAg levels.
2. Viral Supernatants:
   • Clarify by centrifugation to remove debris before testing.

Storage Conditions

• Store all reagents as per the manufacturer’s instructions, typically at 2–8°C.
• Avoid repeated freeze-thaw cycles to maintain reagent integrity.

The QuickTiter™ Hepatitis B Surface Antigen (HBsAg) ELISA Kit is an essential tool for researchers studying HBV infection, vaccine efficacy, and antiviral therapies. Its high sensitivity, specificity, and ease of use make it an ideal choice for detecting and quantifying HBsAg in a variety of research and diagnostic applications.