Human Hemoglobin H (HbH) ELISA Kit

The Human Hemoglobin H (HbH) ELISA Kit is a high-sensitivity, sandwich-based immunoassay designed for the quantitative measurement of Hemoglobin H (HbH) in human biological samples including serum, plasma, urine, cell culture supernatants, tissue lysates, and other body fluids. Hemoglobin H is a key biomarker associated with α-thalassemia and hemolytic anemia, making this kit ideal for research into inherited blood disorders and erythrocyte function.

Key Features

• Assay Type: Sandwich ELISA
• Sample Volume Required: 10 µL
• Detection Wavelength: 450 nm
• Validated Sample Types: Serum, plasma, urine, cell supernatants, tissue homogenates
• Storage: 2–8 °C
• Shelf Life: 6 months
• Assay Format: 96-well microplate (strip format)

Assay Principle

The kit employs a double-antibody sandwich format. The pre-coated microplate wells capture HbH from samples. An HRP-conjugated secondary antibody specific to HbH binds to the captured antigen. Upon addition of TMB substrate, a colorimetric reaction occurs. The resulting color intensity is measured at 450 nm, with signal strength directly proportional to the HbH concentration in the sample. The concentration is determined using a standard curve.

Sample Preparation Guidelines

• Serum: Allow whole blood to clot for 10–20 min at room temperature, then centrifuge at 2,000–3,000 rpm for 20 minutes.
• Plasma: Collect in anticoagulant tubes (EDTA or citrate), incubate 10–20 minutes at room temperature, then centrifuge as above.
• Urine, CSF, Pleural Fluid: Centrifuge at 2,000–3,000 rpm for 20 minutes and collect the supernatant.
• Cell Culture Supernatants: Collect, centrifuge, and use the supernatant directly. For intracellular content, lyse cells via repeated freeze-thaw cycles.
• Tissue Lysates: Homogenize tissues in PBS (pH 7.4), centrifuge at 4 °C, and collect the supernatant for analysis.

Note: Avoid the use of sodium azide (NaN₃) as it inhibits HRP activity. Minimize freeze-thaw cycles to preserve sample integrity.

The Human HbH ELISA Kit offers a reliable, user-friendly platform for the detection and quantification of Hemoglobin H across multiple human sample types. With a small sample volume requirement and high specificity, this kit is a valuable tool for hematological research, particularly in studying α-thalassemia, red blood cell pathologies, and globin chain imbalances. Its compatibility with various biological matrices and straightforward protocol makes it suitable for both basic research and clinical translational studies.