Human Hemoglobin (Hb) ELISA Kit
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Sample Volume: 10ul
Wavelength: 450nm
Assay range: 0.5 μg/ml - 15 μg/ml
Sensitivity: 0.1 ug/ml
Standard: 18 μg/ml
Specifications:
Application | ELISA-Based Assays | ||
Storage Temperature | 2-8°C | ||
Product Type | Elisa Kit | Forms | Kit with Various components |
Product Grade | Molecular Biology | ||
The Human Hemoglobin (Hb) ELISA Kit is a quantitative, sandwich-based enzyme-linked immunosorbent assay designed for the detection and measurement of hemoglobin (Hb) in serum, plasma, cell culture supernatants, tissue homogenates, and other human biological fluids. Hemoglobin is a critical oxygen-transporting protein in erythrocytes, and its levels can serve as a research indicator for anemia, hemolysis, inflammation, and organ injury. This assay provides reliable, sensitive results using only 10 µL of sample per test.
Key Features
- Assay Format: Sandwich ELISA
- Detection Range: 0.5 – 15 μg/mL
- Sensitivity: 0.1 μg/mL
- Sample Volume Required: 10 μL
- Detection Wavelength: 450 nm
- Assay Duration: ~2.5 hours
- Storage Conditions: 2–8 °C
- Shelf Life: 6 months
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Precision:
- Intra-assay CV <10%
- Inter-assay CV <12%
Assay Principle
This kit utilizes a double-antibody sandwich technique. Microplate wells are pre-coated with an anti-Human Hb monoclonal antibody. Upon sample addition, a second HRP-conjugated anti-Hb antibody binds to the captured antigen. A colorimetric reaction is initiated by TMB substrate, producing a blue color that turns yellow upon addition of stop solution. Absorbance is measured at 450 nm, and Hb concentration is calculated using a standard curve.
Kit Contents
Component | Quantity | Storage |
---|---|---|
Microelisa Strip Plate | 96 wells | 2–8 °C |
Hb Standard (18 μg/mL) | 0.5 mL × 1 vial | 2–8 °C |
Standard Diluent | 1.5 mL × 1 vial | 2–8 °C |
Sample Diluent | 6 mL × 1 vial | 2–8 °C |
HRP-Conjugate Reagent | 6 mL × 1 vial | 2–8 °C |
Chromogen Solutions A & B | 6 mL each | 2–8 °C |
Stop Solution | 6 mL × 1 vial | 2–8 °C |
Wash Buffer (30×) | 20 mL × 1 bottle | 2–8 °C |
User Manual | 1 copy | Room Temp |
Closure Plate Membranes | 2 sheets | Room Temp |
Resealable Storage Bag | 1 unit | Room Temp |
Validated Sample Types
- Serum & Plasma (EDTA/citrate anticoagulated)
- Urine, CSF, Pleural Fluid
- Cell Culture Supernatants & Lysates
- Tissue Homogenates
Note: Do not use samples containing sodium azide (NaN₃), which inhibits HRP. Avoid repeated freeze-thaw cycles.
Assay Procedure Summary
- Standard Preparation: Serially dilute from 18 → 1 μg/mL.
- Sample Addition: 40 μL sample diluent + 10 μL sample per well (except blank).
- Incubation: 30 min at 37 °C.
- Wash: 5× using diluted wash buffer.
- Add HRP-Conjugate: 50 μL/well, incubate 30 min at 37 °C.
- Wash Again, then add 50 μL each of Chromogen A & B.
- Color Development: Incubate 15 minutes in the dark.
- Stop Reaction: Add 50 μL stop solution; read OD at 450 nm within 15 minutes.
Performance Characteristics
Parameter | Specification |
---|---|
Detection Range | 0.5 – 15 μg/mL |
Sensitivity | 0.1 μg/mL |
Intra-assay CV | <10% |
Inter-assay CV | <12% |
A log-log standard curve is plotted to determine concentrations. Multiply final values by dilution factor (typically 5) for original concentration.
The Human Hemoglobin (Hb) ELISA Kit (SL2377Hu) provides a reliable, sensitive, and reproducible solution for measuring total hemoglobin in various human biological fluids. Its broad applicability, low detection limit, and minimal sample requirement make it a valuable research tool in hematology, nephrology, toxicology, and clinical pathology. With simple protocols and robust performance, this kit is well-suited for academic, pharmaceutical, and clinical laboratories conducting hemoglobin-related studies.
- Pack Size: For 48 Tests For 96 Tests