Human Anti-Hepatitis B Surface Antibody, Anti-Hbsab Elisa Kit

The Human Anti-Hepatitis B Virus Surface Antibody (HBs-Ab) ELISA Kit is a quantitative enzyme-linked immunosorbent assay (ELISA) designed for the in vitro detection and quantification of HBs-Ab activity in human serum, plasma, cell culture supernatants, and other biological fluids.

This kit utilizes a sandwich ELISA format, offering high sensitivity and specificity through the detection of enzyme-active anti-HBs antibodies using HRP-conjugated antigens and a chromogenic substrate system. The assay is suitable for research applications involving immune response evaluation to Hepatitis B virus surface antigens.

⚠️ For Research Use Only. Not for Diagnostic or Therapeutic Applications.

Key Features

• Sandwich ELISA-based detection
• High sensitivity: 0.5 IU/L
• Quantitative range: 5–100 IU/L
• Reproducible results with CV <10% (intra-assay), <12% (inter-assay)
• Suitable for serum, plasma, cell lysates, and culture media
• Total 96 tests/kit
• Storage: 2–8°C; valid for 6 months

Intended Use

This ELISA kit is intended for the quantitative assessment of enzyme-active Human HBs-Ab in a variety of sample types including:

• Serum
• Plasma (EDTA or citrate-treated)
• Cell culture supernatants
• Tissue homogenates
• Cell lysates

This kit is particularly suitable for monitoring immune response to hepatitis B surface antigens in research studies.

Assay Principle

This kit employs a Sandwich ELISA method:

1. The pre-coated microplate wells contain immobilized Hepatitis B surface antigen (HBsAg).
2. Samples and standards containing HBsAb are added and bind to the coated antigen.
3. A horseradish peroxidase (HRP)-conjugated HBsAg is added to form a sandwich complex with bound HBsAb.
4. After washing to remove unbound components, TMB substrate is added and color develops.
5. The reaction is stopped and optical density is measured at 450 nm.
6. OD values are proportional to the concentration of HBs-Ab in the sample.

 Kit Components (96 Tests)

Sample Preparation Guidelines

• Serum/Plasma: Centrifuge at 2,000–3,000 rpm for 20 minutes. Avoid hemolysis and precipitates.
• Cell culture supernatants: Centrifuge at 2,000–3,000 rpm for 20 minutes.
• Cell lysates: Use PBS (pH 7.2–7.4), subject to repeated freeze-thaw cycles, and centrifuge.
• Tissue homogenates: Homogenize tissue in PBS, centrifuge, and collect supernatant.
• Storage: Store samples at –20°C if not assayed immediately. Avoid repeated freeze-thaw cycles.

Sodium azide (NaN₃) should not be present in samples as it inhibits HRP enzyme activity.

Assay Procedure (Summary)

1. Prepare a standard curve using serial dilutions: 135, 90, 60, 30, 15, and 7.5 IU/L.
2. Add 50 µL of standard/sample + 40 µL sample dilution buffer into each well.
3. Incubate at 37°C for 30 min, then wash 5 times with diluted wash buffer.
4. Add 50 µL of HRP-conjugate reagent, incubate again for 30 min at 37°C.
5. Wash as before and add 50 µL of Chromogen A and B to each well.
6. Incubate in the dark at 37°C for 15 min.
7. Add 50 µL of Stop Solution to terminate the reaction.
8. Read absorbance at 450 nm within 15 minutes.

Performance Characteristics

• Assay Range: 5–100 IU/L
• Sensitivity: 0.5 IU/L
• Precision:
  • Intra-assay CV <10%
  • Inter-assay CV <12%

A standard curve is plotted (log-log scale), and sample concentration is extrapolated by comparing the OD value with standard curve values and applying the appropriate dilution factor.

Storage & Stability

• Storage Temperature: 2–8°C
• Shelf Life: 6 months from date of receipt

Important Notes

• All reagents should be brought to room temperature before use.
• Avoid light exposure during color development (TMB reaction).
• Use fresh pipette tips for every sample to prevent cross-contamination.
• Do not mix reagents from different kits or lots.

The Human HBs-Ab ELISA Kit (Enzyme Activity) offers researchers a reliable, sensitive, and convenient method to evaluate the immune response to hepatitis B virus in human biological samples. With its sandwich ELISA format, excellent precision, and wide assay range, this kit is an ideal tool for vaccine research, immunological studies, and hepatitis B monitoring in experimental settings.